Annals of Tropical Medicine
and Parasitology
THE UNIVERSITY OF LIVERPOOL
Annals
OF
Tropical Medicine and
Parasitology
ISSUED BY THE
Liverpool School of Tropical Medicine
Edited by
Professor J. W. W. STEPHENS, M.D.Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
_yorke, m.d.
Annals of Tropical Medicine and Parasitology
Vol. VIII, p. 585, lines 8-10. The authors, Drs. Breinl and
Young, wish to amend ‘Any .... dryness;’ to read thus:—‘Any
lead sulphide was filtered off, washed with sulphuretted hydrogen
‘water, dissolved in dilute nitric acid, and the excess of acid
‘ removed by repeatedly evaporating to dryness
With Frontispiece , thirty-nine plates , twenty-pve figures in text, menty-jour
charts, and three maps
LIVERPOOL:
AT THE UNIVERSITY PRESS, 57 ASHTON STREET
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool .
CONTENTS
No. i. March 18, 1915
Bayon, H. page
Leprosy : A Perspective of the Results of Experimental Study of the Disease.
Plates I-VI . 1
Young, W. J.
The Metabolism of White Races living in the Tropics. I.—The Protein
Metabolism . 91
Ward, Henry B. ; and ITirscii, Edwin F.
The Species of Paragonimus and their Differentiation. Plates YII-XI ... 109
ScmvETZ, Dr. J.
Preliminary Notes on the Mosquitos of Kabinda (Lomami), Belgian Congo... 163
Stephens, J. W. W.
On the Peculiar Morphological Appearances of a Malaria Parasite. Plate XII 169
Carter, Henry F.
On Some Previously Undcscribed Tabanidac from Africa. Plate XIII . . 173
Seidelin, Harald.
Experiments with Salvarsan-Copper in Trypanosomiasis . 197
Stephens, J. W. W. ; and Stott, W.
Studies in Blackwater Fever. III.—The Relationship of Quinine to Blackwatcr
Fever . 201
CONTENTS
No. 2. June 30, 1915
Brf.inl, Anton. pace
Gangosa in New Guinea and its Etiology. Plates XIV-XVII . 213
Priestley, Henry.
TheiUria tachyglossi (n.sp.) a Blood Parasite of Tachyglossus aculcatus. Plate
XVIII. 233
Scott, H. Harold.
An Investigation into the Causes of the Prevalence of Enteric Fever in Kingston,
Jamaica ; with Special Reference to the Question of Unrecognised Carriers 239
CONTENTS No. 2 — continued
Rrkinl, An ion. page
On the Occurrence and Prevalence of Diseases in British New Guinea. Plates
XIX-XXVI . 285
Fantham, H. B.
Inject Flagellates and the Evolution of Disease, with Remarks on the Importance
of Comparative Methods in the Study of Protozoology ... ... ... 335
CONTENTS
No. 3. July 31, 1915
PACK
Yorke, Warrington ; and Blacklock, B.
Notes on the Bionomics of Glossina palpalis in Sierra Leone, with Special
Reference to its Pupal Habitats. Plates XXVII-XXXIII . 349
Yorke, Warrington; and Blacklock, B.
Food of Glossina palpalis in the Cape Lighthouse Peninsula, Sierra Leone ... 363
Fraser, Henry ; and Fletcher, W.
The Cultivation of the Leprosy Bacillus. 381
Yorke, Warrington ; and Blacklock, B.
The Reservoir of the Human Trypanosome in Sierra Leone. 383
Fantham, H. B.
Spirocbaeta hronchialis , Castcllani, 1907, together with Remarks on the Spiro-
chaetes of the Human Mouth. Plate XXXIV . . 391
Yorke, Warrington ; and Blacklock, B.
Notes on Certain Animal Parasites of Domestic Stock in Sierra I,cone ... 413
Davey, J. B.
The Etiology of Juxta-articular Subcutaneous Nodules ... ... ... 421
Yorke, Warrington ; and Blacklock, B.
Ankylostomiasis in Dogs in Sierra Leone. 425
Stephens, J. W. W.
Studies in Blackwatcr Fever. IV.—Note on a Case of Quartan Malaria
associated with Blackwater Fever . 429
CONTENTS
No. 4. December 30, 1915
PAGE
Macfie, J. W. Scott
Nuclear Variations of the Ncutrophilc Leucocytes (Arnetli Counts) in Malaria
and Yellow Fever . 435
Macfie, J. VV. Scott
Babesiasis and Trypanosomiasis at Accra, Gold Coast, West Africa. Plates
XXXV, XXXVI . 457
Breinl, A. ; and Priestley, H.
Differential Counts and the Neutrophile Blood-Picture of Natives-—Adults
and Children—of New Guinea . 495
Macfie, J. W. Scott
A Case of Dysentery in a Monkey, in which Amoebae and Spirochaetcs were
found. Plate XXXVII . 507
Schwetz, Dr. J.
Preliminary Note on the General Distribution of Glossina pal pahs , Rob-Dcsv.,
in the District of Lomami, Belgian Congo. 513
Macfie, J. W. Scott
A Note on a Trypanosome of the Black Rat (Epimys rattus). Plate XXXVIII 527
Bayon, H.
The Artificial Cultivation of Hansen’s ‘ Bacillus’ . 535
Stephens, J. W. W.
Studies in Blackwater Fever. V.—The Duration of Haemoglobinuria ... 539
Fantham, H. B.; and Porter, Annie
Some Experimental Researches on Induced Herpetomoniasis in Birds.
Plate XXXIX. 5+3
Lloyd, Ll.
On the Association of VVarthog and the Nkufu Tick ( Ornithodorus moubata) ... 559
Porter, Annie
On Anaplasma-like Bodies in the Blood of Vertebrates ...
561
Volume IX
March, 1915
No. 1
ANNALS
# OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor WARRINGTON YORKE, M.D.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool .
C. Tinting < 5 r* Co., Ltd.
Printers to the University Press of Liverpool
53 Victoria Street
THE INCORPORATED
LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Founded by Sir ALFRED LEWIS JONES, K.C.M.G.
{Affiliated with the University of Liverpool)
Hon. President: H.R.H. Princess Christian of Schleswig-Holstein.
Chairman : MR. F. C. DANSON.
Vice-Chairman: Professor R. CATON.
Hon. Vice-Presidents: The Duke of Northumberland, K.G.
The Earl of Derby, K.G.
Earl Cromer, G.C.B.
Viscount Milner, G.C.B.
Lord Pirrie, K.P.
Sir Edward Merewether, K.C.V.O.
Sir Owen Philipps, K.C.M.G.
Sir William H. Lever, Bart.
Mr. O. Harrison Williams
COMMITTEE
Mr. H. J. Read, C.M.G.
Vice-Chancellor Sir Alfred Dale
Sir W. B. Bowring, Bart.
Professor R. CATON
Professor W. HERDMAN, F.R.S.
Professor J. M. Beattie
Dr. C. J. Macalister
M r. G. P. Newbolt
M r. C. Booth (Jun.)
Mr. T. F. Harrison
Mr. A. R. Marshall
Mr. W. Roberts
Mr. J. W. Alsop
Mr. G. Brocklehurst
Mr. C. Livingston
Colonial Office
University of Liverpool
■ Council of University of Liverpool
| Senate of University of Liverpool
Royal Southern Hospital
Steamship Owners’ Association
Shipowners’ Association
Mr. J. W. W. Danson
Professor E. W. HOPE
Mr. R. Rankin
Professor E. GLYNN
Mr. E. G. Buckley
Mr. A. L. Rea, Hon. Treasurer
Mr. A. H. MILNE, C.M.G., Secretary.
B io Excharge Buildings, Liverpool
Staff, 1915
1. At the University of Liverpool
Professors - - JOHN WILLIAM WATSON STEPHENS, M.D.,
Cantab., D.P.H. Sir Alfred Jones Professor of
Tropical Medicine
ROBERT NEW STEAD, E.R.S., M.Sc., A.L.S.,
F.E.S., Dutton Memorial Professot of Entomology'
LiF.rT.-Coi., Sir RONALD ROSS. K.C.B., F.R.S', -
LL.D., E.R.C.S., D.P.H., M.D., D.Sc. Nobel
Laureate 1902. (Indian Medical Service, retired>.
Professor of Tropical Sanitation
WARRINGTON YORKE, M.D.. If alter Myers
Professor of Parasitology
Lecturers - Prof. E. W. HOPE, M.D., D.Sc., Lecturer or
Municipal Sanitation
HENRY FRANCIS CARTER, S.E.A.C. Dim..
F.E.S., Lecturer on Entomology
HAROLD BENJAMIN FANTHAM, M.A., D.Sc.,
Lecturer on Parasitology
WILLIAM THOMAS PROUT, M.B , C.M.G.,
Lecturer on Tropical Sanitation
HAROLD WOLFERSTAN THOMAS, M.D.,
C.M.
Honorary Lecturer - Major JOSEPH FITZGERALD BLOOD, M.D..
M.Ch. (Indian Medical Service, retired)
Honorary Statistician WALTER STOTT
2. At the Research Laboratories
Director - - - B. BLACKLOCK, M.D., D.P.H.
3. At the Royal Infirmary, Liverpool
Physician - - JOHN WILLIAM WATSON STEPHENS, M I).,
Assistant to the Cantab., D.P.H.
Physician - WILLIAM THOMAS PROUT, M.B , C.M.G.
4. At the Royal Southern Hospital, Liverpool
Phys icians
Surgeon
- CHARLES JOHN MAC A LISTER, M.D.,
F.R.C.P.
JOHN LLOYD ROBERTS. M.D.. M.R.C.P.
- GEORGE PALMERSTON NEWBOLT, M B.,
F.R.C.S.
Hon. Tropical
Pathologist
JOHN WILLIAM WATSON STEPHENS, M.D..
Cantab., D.P.H.
5. At the Yellow Fever Research Laboratory of the School,
Manaos
Director - - HAROLD WOLFERSTAN THOMAS, M.D.,
C.M.
Laboratory
JOHNSTON LABORATORY, UNIVERSITY OF LIVERPOOL
Hospitals:
ROYAL INFIRMARY AND ROYAL SOUTHERN HOSPITAL,
LIVERPOOL
Secretary's Office:
B 10, EXCHANGE BUILDINGS, LIVERPOOL
NOTICE
The following courses of instruction will be given by the
Liverpool School of Tropical Medicine during 1916 :—
Full Course begins 6 January. Advanced Course begins 1 June.
Diploma Examination, 3 April. Certificate Examination, 30 June.
Full Course begins 15 September.
Diploma Examination, 11 December.
These dates are subject to revision.
The full Course of Instruction is open to all qualified medical men,
and the Examination to all students who have taken out this full
course.
Fee for the full Course of Instruction—Thirteen Guineas.
Fee for the Diploma Examination—Five Guineas.
Fee for the Short Course of Instruction—Four Guineas.
Fee for the use of a School microscope during one term—Ten
shillings and sixpence.
For prospectus and further information, application should be made
to the Dean of the Medical Faculty, University of Liverpool.
The following have obtained the Diploma in Tropical Medicine of
the University of Liverpool: —
Diploma in Tropical Medicine
Date 0 /
Difloma
1904 Augustine, Henry Joshua
1904 Bennett, Arthur King
1904 Bruce, William James
1904 Byrne, John Scott
1904 Clayton, Thomas Morrison
1904 Dalziel, John McEwen
1904 Dee, Peter
1904 Greenidge, Oliver Campbell
1904 Hehir, Patrick
1904 Khan, Saiduzzafor
1904 Laurie, Robert
1904 Maclurkin, Alfred Robert
1904 McConnell, Robert Ernest
1904 Nicholson, James Edward
1904 Philipson, Nicholas
1904 Sharman, Eric Harding
1904 Thomson, Frank Wyville
1904 Walker, George Francis Clegg
1905 Anderson, Catherine F.lmslie
1905 Brown, Alexander
1905 Caldwell, Thomas Cathcart
1905 Critien, Attilio
1905 Hooton, Alfred
1905 Hudson, Charles Tilson
1905 Illington, Edmund Moritz
Date oj
Diploma
1905 Macfarlane, Robert Maxwell
1905 Maddock, Edward Cecil Gordon
1905 Moore, James Jackson
1905 Nightingale, Samuel Shore
1905 Radcliffe, Percy Alexander Hurst
1905 Young, John Cameron .
1906 Adie, Joseph Rosamond
1906 Arnold, Frank Arthur
1906 Bate, John Brabant
1906 Bennetts, Harold Graves
1906 Carter, Robert Markham
1906 Chisholm, James Alexander
1906 Clements, Robert William
1906 Dundas, James
1906 Faichnie, Norman
1906 Jeffreys, Herbert Castelman
1906 Mackenzie, Donald Francis
1906 Pailthorpe, Mary Elizabeth
1906 Palmer, Harold Thornbury
1906 Pearse, Albert
1906 Sampey, Alexander William
1906 Smithson, Arthur Ernest
iqo6 Taylor, Joseph van Someron
1906 Taylor, William Irwin
1906 Tynan, Edward Joseph
Date of
Difloma
1906 Watson, Cecil Francis
1906 Willcocks, Roger Durant
1906 Williamson, George Alexander
1907 Allan, Alexander Smith
1907 Allwood, James Aldred
1907 Bond, Ashton
1907 Branch, Stanley
1907 Collinson, Walter Julius
1907 Davey, John Bernard
1907 Donaldson, Anson Scott
1907 Fell, Matthew Ilenry Gregson
1907 Gann, Thomas William Francis
1907 Graham, James Drummond
1907 Hiscock, Robert Carroll
1907 Keane, Joseph Gerald
1907 Kennan, Richard Henry
1907 Kenrick, William Hamilton
1907 Le Fanu, George Ernest Hugh
1907 Mackey, Charles
1907 Maddox, Ralph Henry
1907 McCarthy, John McDonald
1907 Raikes, Cuthbert Taunton
1907 Ryan, Joseph Charles
1907 Vallance, Hugh
1908 Caverhill, Austin Mack
1908 Crawford, Gilbert Stewart
1908 Dalai, Kaikhusroo Rustomji
1908 Dansey-Browning, George
1908 Davidson, James
1908 Dickson, John Rhodes
1908 Dowdall, Arthur Melville
1908 Glover, Henry Joseph
1908 Greaves, Francis Wood
1908 Goodbody, Cecil Maurice
1908 Harrison, James Herbert Hugh
1908 Joshi, Lemuel Lucas
1908 Le Fanu, Cecil Vivian
1908 Luethgen, Carl Wilhelm Ludwig
1908 Mama, Jainshed Byramji
1908 McCay, Frederick William
1908 McLellan, Samuel Wilson
1908 Pearce, Charles Ross
1908 Schoorel, Alexander Frederik
1908 Smith, John Macgregor
1908 Stewart, George Edward
1908 Tate, Gerald William
1008 Whyte, Robert
1909 Abercrombie, Rudolph George
1909 Allin, John Richard Percy
1909 Armstrong, Edward Randolph
1909 Barrow, Harold Percy Waller
1909 Beatty, Guy
1909 Carr-White, Percy
1909 Chevallier, Claude Lionel
1909 Clark, William Scott
1909 Cope, Ricardo
1909 Fleming, William
1909 Hanschell, Hother McCormick
1909 Hayward, William Davey
1909 Henry, Sydney Alexander
1909 Innes, Francis Alexander
Date of
Diploma
1909 Jackson, Arthur Frame
1909 Kaka, Sorabji Manekji
1909 McCabe-Dalfas, Alfred Alexander
Donald
1909 Meldrum, William Percy
1909 Murphy, John Cullinan
1909 Samuel. Mysore Gnananandaraju
1909 Shroff, Kawasjee Bvramjee
1909 Thornely, Michael Harris
1909 Turkhud, Violet Ackroyd
1909 Webb, William Spinks
1909 Yen, Fu-Chun
1910 Brabazon, Edward
1910 Castellino, Louis
1910 Caulcrick, James Akilade
1910 Dowden, Richard
1910 Haigh, William Edwin
1910 Hamilton, Henry Fleming
1910 Hefferman, William St. Michael
1910 Hip well, Abraham
1910 Homer, Jonathan
1910 Houston, William Mitchell
1910 James, William Robert Wallace
1910 Johnstone, David Patrick
1910 Korke, Vishnu Tatyaji
1910 Macdonald, Angus Graham
1910 Macfie, John Wm, Scott
1910 Manuk, Mack Walter
1910 Murison, Cecil Charles
1910 Nanavati, Kishavlal Balabhai
1910 Nauss, Ralph Welty
1910 Oakley, Philip Douglas
1910 Pratt, Ishmael Charles
1910 Sabastian, Thiruchelvam
1910 Shaw, Hugh Thomas
1910 Sieger, Edward Louis
1910 Sousa, Pascal John de
1910 Souza, Antonio Bernardo de
1910 Waterhouse, John Howard
1910 White, Maurice Forbes
1911 Blacklock, Breadalbane
1911 Brown, Frederick Forrest
1911 Chand, Diwan Jai
1911 Holmes, John Morgan
1911 levers, Charles Langley
1911 lies, Charles Cochrane
1911 Ingram, Alexander
1911 Kirkwood, Thomas
1911 Knowles, Benjamin
1911 Liddle, George Marcus Berkeley
1911 Lomas, Emanuel Kenworthy
1911 Mackarell, William Wright
1911 MacKnight, Dundas Simpson
1911 Mascarenhas, Joseph Victor
1911 Murray, Ronald Roderick
1911 Oluwole, Akidiva Ladapo
1911 Rao, Koka Ahobala
1911 Sinton, John Alexander
19 n Tarapurvalla, Byramji Shavakshah
1911 Taylor, John Archibald
1911 Woods, William Medlicott
Date of
Diploma
1912 Aeria, Joseph Reginald
1912 Anderson, Edmund Litchfield
1912 Borle, James
1912 Bowie, John Tait
1912 Brassey, Laurence Percival
19 1 2 Christie, David
1912 Dillon, Henry de Courcy
1912 Dunn, Lillie Eleanor
1912 Hardwicke, Charles
1912 Jagose, Jamshed Rustomji
1912 Kochhar, Mela Ram
1912 McGusty, Victor William Tighe
1912 Milne, Arthur James
1912 Mitra, Manmatha Nath
1912 Myles, Charles Duncan
1912 Pelly, Huntly Nevins
1912 Prasad, Bindeshwari
1912 Prentice, George
1912 Ross, Frank
1912 Russell, Alexander James
Hutchison
1912 Ruthven, Morton Wood
1912 Sandilands, John
1912 Seddon, Harold
1912 Smalley, James
1912 Strickland, Percy Charles
Hutchison
1912 Watson, William Russel
1913 Austin, Charles Miller
1913 Banker, Shiavux Sorabji
1913 Becker, Johann Gerhard us
1913 Carrasco, Milton
1913 Clark, James McKillican
1913 Forsyth, Charles
1913 Grahame, Malcolm Claude Russell
1913 Grieve, Kelburne King
1913 Hargreaves, Alfred Ridley
1913 Hepper, Evelyn Charles
1913 Hiranand, Pandit
1913 Jackson, Oswald Egbert
1913 IChaw, Ignatius Oo Kek
1913 MacKelvie, Maxwell
1913 MacKinnon, John MacPhail
1913 Macmillan, Robert James Alan
Date of
Diplotna
1913 Mouat-Biggs, Charles Edward
Forbes
1913 Noronha, John Carmel
1913 O’Connor, Edward
1913 Olubomi-Beckley, Emanuel
1913 Pestonji, Ardeshir Behramshah
1913 Puttanna, Dodballapur Sivappa
1913 Reford, John Hope
1913 Smith, Edward Arthur
1913 Stewart, Samuel Dudley
1913 Walker, Frederick Dearden
1913 Wilbe, Ernest Edward
1913 Wilson, Hubert Francis
1913 Yin, Ulg Ba
1913 Young, William Alexander
1914 Arculli, Hassan el
1914 Chohan,Noormahomed Kasembha
1914 Connell, Harry Bertram
1914 Gerrard, Herbert Shaw
1914 Gimi, Hirji Dorabji
1914 Gwynne, Joseph Robert
1914 Hodkinsou, Samuel Paterson
1914 Jackson, Arthur Ivan
1914 Kaushash, Ram Chander
1914 Kelsall, Charles
1914 Luanco y Cuenca, Maximino
1914 Misbah, Abdul-Ghani Naguib
1914 Naidu, Bangalore Pasupulati
Balakrislma
1914 Rowe, John Joseph Stephen
1914 Roy, Raghu Nath
1914 Sbiveshwarkar, Ramchandra
Vishnu
1914 Sur, Sachindra Nath
1914 Talati, Dadabhai Cursedji
1914 Wilkinson, Arthur Geden
1914 Wright, Ernest Jenner
1915 Lobo, John Francis
1915 Madbok, Gopal Dass
1915 Pearson, George Howorth
1915 Swami, Karumuri Virabhadra
1915 Wood, John
ANNALS OF TROPICAL MEDICINE
AND PARASITOLOGY
EDITORIAL NOTICE
Articles for publication are accepted, if found suitable, from any
source and will be understood to be offered alone to these Annals.
They should be typewritten and addressed to: —The Editorial
Secretary, School of Tropical Medicine, The University, Liverpool.
Illustrations for text figures should be drawn clearly and firmly
in Indian ink, if possible, on Bristol board.
All lettering, names or legends on text-figures, charts or maps
should be printed sufficiently large to allow of clear legibility on
reduction if necessary.
Plates and illustrations should be accompanied by short explana¬
tions.
References to authors in the text must be made in the following
way:—'According to Smith (1900) the spleen is enlarged, but
Robinson (1914) says the reverse.’ The references should be
collected in alphabetical order of authors’ surnames at the end of
the paper, and arranged in the following way : —
Robinson. S. (1914). ‘The spleen in malaria.' Annah of Nosology,
Vol. XX. pp. 20-25.
Smith, J. (1900). ‘ Enlargement of the spleen in malaria.’ Journal of
Pathometry , Vol. I, pp. 1-20.
Fifty reprints are supplied to each author, free of charge.
Additional copies (up to 200) can be supplied at cost price.
Subscription: £\ 2s. 6d. per volume, post free, payable in
advance to the Secretary, Liverpool School of Tropical Medicine,
B 10 Exchange Buildings, Liverpool. Correspondence concerning
advertisements should also be addressed to the Secretary.
LEPROSY: A PERSPECTIVE OF THE
RESULTS OF EXPERIMENTAL STUDY
OF THE DISEASE
BY
H. BAYON, M.D.,
RESEARCH BACTERIOLOGIST (LEPROSY), GOVERNMENT OF THE UNION OF SOUTH AFRICA,
AT ROBBEN ISLAND
(Received for publication 18 August , 1914)
Plates I—VI
CONTENTS
I—Introduction.
II —The Clinical Nosology or Leprosy .
III— Microscopical Features or Leprous Lesions .
IV — Morphology and Staining Properties or Hansen's 4 Bacillus *
Analogy with the Mycobacterium tuberculosis .
V —Artificial Cultivation or the Micro-organism or Leprosy
VI— Experimental Transmission to Animals.
Rat-Leprosy and its Relation to the Human Disease .
VII —Serological Tests and Reactions in Leprosy.
VIII —Communicability and Contagion.
Is the transmission through an intermediate arthropod probable ?
IX— Early and Difperential Diagnosis. Prognosis .
X— Treatment, Specific and Palliative .
XI —Prevention. Home and Asylum Segregation. Settlements
XII —Discussion or Results and Conclusions.
Explanation or Plates .
4
9
18
20
23
3 1
40
47
55
6o
64
70
73
77
8 o
I. INTRODUCTION
Leprosy is the name applied to all clinical manifestations
produced by an infection with the micro-organism commonly known
as ‘Hansen’s bacillus’ {Mycobacterium leprae).
It is a disease that has been recognised and feared for centuries,
possibly already as far back as 2400 B.C., or in any case about
1300 B.C. (Ebers papyrus). Leprosy as described in Leviticus does
2
not, however, correspond exactly to the symptoms we know in more
modern times. It should, moreover, not be forgotten that the word
' leprosy ’ is but an approximate rendering of the expression
‘zaraath’ in the original Hebrew.
Not only is leprosy the ‘ most ancient disease,’ but it is also a
widely spread, cosmopolitan scourge, which nearly rivals con¬
sumption, syphilis and cancer, if not in numbers, at least in the
variety of climatic conditions under which it can flourish and
increase. Though at the present day it is found, to any con¬
siderable extent, practically only in tropical and sub-tropical
countries, it is known that numerous lepers existed in France,
Germany, Great Britain, and other countries of Europe throughout
the Middle Ages. Opinions differ whether it was imported into
Europe from the East through the returning Crusaders. The
disease still lingers in Turkey, Russia, the Balkan States,
Norway, Spain, Portugal, parts of Italy, etc., in the form of small
endemic foci. However, the ruthless individual and, in some cases,
collective segregation in the thirteenth and fourteenth centuries,
aided no doubt by the improving hygienic conditions of all classes,
succeeded in bringing about a practically complete extinction of
leprosy in Middle Europe and Great Britain. In addition to this,
various plague epidemics also helped in sweeping to an early grave,
in a somewhat selective fashion, the vagrant and pauper lepers
sooner and quicker than individuals belonging to better situated
classes.
The slow if intermittent course of the disease, the extensive
mutilations and repulsive features, which are an outstanding
symptom of its more advanced stages, have rendered leprosy a
subject of terror and horror from time immemorial. That it is
contagious has been known for centuries, but it is no doubt mainly
due to aesthetic reasons that the fear it engendered was quite out
of proportion to the degree in which it is communicable, under
usual circumstances, from the diseased to the healthy.
Leprosy presents numerous peculiarities in its pathognomonic
features. Not only is its infectivity extremely variable under slightly
dissimilar or even identical circumstances, but its period of incuba¬
tion varies also within very wide limits, anything between one year
and twenty-five or thirty. The ‘causa prima morbi’ has been
3
known for years and years, and yet the fundamental bases of all
bacteriological research, viz., artificial cultivation of the micro¬
organism and the experimental transmission of the disease, have
presented insurmountable difficulties up to the present day.
Indeed, these bases are still matters under discussion, and in any
case successful results can only be achieved after numerous,
repeated, prolonged, pertinacious attempts. The large numbers of
bacteria which may be present in single leprotic lesions are certainly
not surpassed in any other disease, with the possible exception of
bubonic plague.
An excellent instance showing the influence that bacteriology
exercises on clinical medicine is to be found in the effects resulting
from the discovery of the ‘ bacillus ’ of leprosy by Armauer Hansen
in 1872. It is true that definite knowledge of the morphology of
the micro-organism was only obtained after Neisser had succeeded
in demonstrating it by means of an ‘acid-fast' stain in 1879.
However, from the moment the bacterial origin of the disease was
recognised, the communicability of leprosy became scientifically
established, and all doubts in that connection could only be the
outcome of an incomplete or lacking comprehension of the applica¬
tion of the laws of biology to practical medicine.
On the other hand the extreme difficulty encountered in
transmitting the disease to animals, the repeated lack of success
attending the inoculation of leprous material into human beings
(with one exception: Aming), have allowed much doubt to arise
regarding the exact fashion in which leprosy is transmitted. Even
the purely bacteriological observation of the fact that apparently
the ‘bacilli’ could not be artificially cultivated, and isolated from
the lesions they produced, has seriously hampered the therapeutic
outlook.
Leprosy slowly and surely became the disease of negations; it
could not be experimentally transmitted, its bacterium could not be
isolated in artificial culture, and so to complete the list, leprosy
could not be treated, let alone arrested, and no hope of cure could
be entertained.
It is my intention to show whether recent experimental work
allows, enables, or justifies us in taking a more active and positive
view of the situation.
II. THE CLINICAL NOSOLOGY OF LEPROSY
Two clinical types of the disease are generally distinguished
and no more. Such views are partly owing to the influence of
Hansen, who considered that it was not necessary to make further
sub-divisions because practically all lepers presented features at one
time or another of their existence (that is, if they lived long
enough) which allowed them to be classed either as cases of
(i) nodular (hypertrophic) leprosy, or (ii) maculo-anaesthetic, other¬
wise called atrophic, smooth leprosy.
From a purely clinical point of view this classification holds
good, however, only within certain limits. It is a commonly
accepted rule of nosology that bedside or clinical nomenclature
does not take into consideration so much the eventual course of a
disease, as its etiology and the symptoms and appearances which,
however fleeting and changeable, can be brought under a definite
scheme of relative incidental constancy. The classification used in
relation to recognised clinical types of tuberculosis, syphilis, and
other complaints, which often run a chronic course and vary largely
in their symptomatology, seems clearly to show which are the
determining factors ruling the designation of clinical entities.
It is no doubt quite correct, from a general point of view, to
state, as Hansen did, that a ‘mixed’ leper (i.e., a patient who from
the very beginning shows features of both the nodular and maculo-
anaesthetic type) who lives long enough and undergoes a natural
process of elimination of the great majority of the acid-fast
‘ bacilli ’ from his lesions, will eventually be classifiable as a purely
anaesthetic leper. Many of these instances of * lepra mixta,’
however, do not reach this ultimate stage, because death steps in
before completion of the process of evolution into a different type,
and in any case the primary condition may last for many years.
Without unnecessarily labouring this point any further, I
believe that there is quite sufficient justification in considering every
single case of leprosy as classifiable on the basis of the clinical and
bacterioscopic features at the time of examination, irrespective of
the expected course of the disease later on. In other words, the
clinical diagnosis is to be considered of a temporary nature, as far
as the naked eye appearances (and to a certain extent also the
bacterioscopic results) are concerned. The diagnosis leprosy is the
5
only unchangeable feature, though it may need modification by the
adjectives ‘ arrested ’ or possibly even ‘ cured.’
Leaving aside sub-divisions founded on partial or purely
localised symptoms, I believe that on examination of any great
number of lepers, in any part of the world, more or less numerous
specimens of the following varieties or even distinct types of the
disease may be distinguished.
1. Nodular type with more or less marked or diffuse tuberosities
which generally are most developed on the face, but of course can
also affect any other part of the body, including (though somewhat
rarely) the scalp, the soles of the feet, the tongue, and the palms
of the hands. Leprous nodules should contain numerous acid-fast
micro-organisms, and in certain instances may be located as loose
or movable small lumps which can be shelled out from under the
skin.
Nodular leprosy may give rise to visceral lesions, which are
remarkable for the enormous quantity of acid-fast ‘ bacilli ’ that
can be detected accumulated in the tissues of the spleen, liver,
testicles, lymphatic glands and lungs.
2. Maculo-anaesthetic type with more or less numerous spots
which have a slight predilection for the trunk (chest, back, buttocks)
but are certainly also very frequently present on the limbs. The
spots in advanced cases may show a greater or lesser amount of
anaesthesia. It should be noted that often the anaesthesia is
definitely localized in portions not showing any maculae. In many
cases, however, the anaesthesia is by no means complete, and should
more correctly be termed paraesthesia. The patients cannot
distinguish hot and cold, sharp or blunt in the affected areas, but
are still sensitive to touch. In early stages the maculae may present
a ringed appearance; the centre then may be normal in relation to
sense of touch, etc., whilst the outer ring, if anything, is slightly
hyperaesthetic.
The maculae appear to be the result of the localisation (and
possibly disintegration) of a small number of casual micro¬
organisms in the skin. The anaesthetic areas are due to the
‘ bacilli ’ invading the sheath of the nerves, causing a proliferation
of connective tissue, which brings about a slow degeneration of the
corresponding nerve fibres. In advanced cases the bacilli invade
6
the ganglia of the spinal cord, hence the practical impossibility,
under certain circumstances, of distinguishing leprosy clinically
from syringomyelia.
5. Mixed or combined leprosy, that is, cases where from the
very earliest noticeable onset of the disease, and for years after¬
wards, definite areas of paraesthesia (insensibility to heat and cold,
incapacity of distinguishing blunt from sharp) and anaesthesia can
be made out, accompanied by maculae and small nodules teeming
with acid-fast bacteria.
An excellent description of this type has been given by Gluck,
who called it ‘ lepra tubero-anaesthetica.’
These three main types can be sub-divided into the following
relatively rare varieties and singular forms:
1. Lupoid or serpiginous leprosy. Among the 2,000 lepers I
have had under observation in South Africa I have only met with
the disease three times. Another instance of the variety was seen
in London. It is recognisable as leprosy by the presence of nerve-
lesions, but acid-fast micro-organisms are very scarce indeed in any
of the superficial patches. The expression ‘ patches ’ is used
because the skin-lesions are much more raised and thickened than
mere ‘ maculae,’ and yet do not deserve the appellation of nodules,
being too flat and extensive. They spread by forming peculiar
confluent configurations, which can be best recognised by referring
to PI. V, fig. 1.
The patches are a peculiar coppery colour, and do not show any
marked tendency to break down and ulcerate, which in addition to
other peculiarities distinguishes them from lupus.
At times single patches whose colour, texture and general
appearance corresponds closely to that of these serpiginous lesions,
are noticeable in cases of leprosy, whose remaining clinical features
differ in no other fashion from the usual nodular type.
In other instances the patches may not be confluent and
serpiginous, but are distributed unevenly all over the body, varying
in size from the diameter of a threepenny-bit to the extent of a
man’s hand or more. They may appear relatively rapidly, within
a fortnight or so, and are accompanied by localised paraesthesia.
In such case the diagnosis usually presents considerable difficulty,
because acid-fast bacteria may be practically absent in the lesions
and nasal mucosa.
7
Jadassohn has described somewhat similar varieties of leprosy
as the ‘ tuberkulid ’ type, in view of the resemblance which micro¬
scopical specimens of the lesions bear to tuberculosis of the skin.
2. Many cases of maculo-anaesthetic leprosy begin with
paraesthesia, numbness, and a peculiar ‘ leathery ‘ feeling, usually
of the ring and little finger of one hand, or tingling sensations with
small areas of definite anaesthesia may be present in a hand or
foot. Such conditions can last for a considerable time, as much as
three or more years, before definite maculae make their appearance.
Some cases, after a prolonged premonitory stage of this nature, may
develop nodules and finish as definite hypertropic leprosy.
Maculo-anaesthetic lepers may heal in such a fashion as to allow
hardly any traces of their spots being visible to the naked eye; the
trained observer may just be able to make out a simple configurated
discoloration or slight livido. Needless to say, definite areas of
anaesthesia are usually easily made out in these patients. As long
as these conditions last it appears quite correct to classify them as
simple paraesthetic or anaesthetic leprosy.
3. Under certain circumstances maculae or spots may appear
before any nerve symptoms are traceable. Even to the experienced
clinician the leprotic nature of the spots may appear doubtful,
other conclusive symptoms being absent and possibly no history of
contact being available, though the latter has but a relative value.
In these rare occasions only prolonged observation, aided and
controlled by the microscope, can afford us certainty in diagnosis.
Eventually, in the course of time, a definite maculo-anaesthetic
stage is reached, but during this initial period the condition is one
of simple macular leprosy.
When dealing with dark-skinned races great care must be taken
not to mistake for macular leprosy one of the numerous tineae which
have been so ably classified and investigated by Castellani.
4. In some instances leprotic symptoms are seen which are
somewhat too prominent to deserve still the name of ‘ maculae ’ or
spots. The edges are considerably more raised than is usually the
case in smooth leprous lesions, owing to active proliferation of the
superficial dermal tissue. The periphery is studded with numerous
minute papulae, which have a somewhat vesicular appearance. The
central portion may or may not participate in this process of
granulation, but a certain degree of desquamation is generally
8
present. In any ease the appearance of the inner area is angrier and
more ‘ active-looking ’ than what is seen in the usual run of maculae,
though parts of the surface do not usually participate to such a
marked degree in the process of granulation and eruption notice¬
able at the margin.
It may be objected that practically all maculae of lepers go
through periods of increased activity or exacerbation, during which
they may show some of the appearances described. However, the
case illustrated in PI. V, fig. 2, has remained in this condition
for over three years, and moreover, the localisation of the lesion
is somewhat peculiar. In addition, the maculae of many lepers
never reach the condition of marginate proliferation shown in the
photograph.
Of course this whole condition is but the result of an intensified
tissue reaction, such as generally takes place to a much lesser
degree in most maculae. The diagnosis, however, may still
present difficulties, because, as in the instance illustrated, other
definite leprotic symptoms, such as anaesthesia, may be absent or
cannot be detected at the time of examination. I should like to
draw attention to the existence of these intensified ‘maculae*
without insisting on their separate classification.
5. Diffuse or hyperaemic leprosy is also scarcely a separate
variety, but simply an initial or premonitory stage of nodular
leprosy. It consists in a peculiar diffuse, puffy swelling of the
eyebrows, the lobes of the ears and the alae nasi, which in addition
takes on a peculiar pinkish-bluish hue. This condition may last for
years and imperceptibly merge into the well-known condition of
leprosy generally known as ‘ facies leonina.’ In rare instances the
disease may not make further progress, but eventually improves and
leaves, as the only trace, a peculiar wrinkled and dry appearance of
the skin of the face. I have so far not found any of such arrested
cases without smaller or larger areas of anaesthesia.
These three types and five varieties or peculiar diversities from
the usual clinical appearances of leprosy appear to me to cover all
forms of leprosy, as far as I have been able to observe them. In
addition, there are numerous combinations which can result through
atypical and transitory appearances of the disease mixing in varying
proportions.
9
Bullae, ulcers, suppurations and necrosis may considerably alter
and add complexity to the clinical features of the symptomatology
of leprosy. They should, however, be considered as complications
of the primary infection; moreover, they are too variable and
inconstant to allow definite systematization.
Careful description of the majority of these symptoms is also
given by Abraham (1910) where special stress is laid on the fact,
and rightly too, that the diagnosis is in many cases but the
description of a stage of the disease.
These various sub-divisions have not been made to complicate
further a diagnosis which under usual circumstances is already
sufficiently difficult, but on the contrary to induce a careful observa¬
tion of the various phenomena of a disease whose correct, exact, and
above all early recognition is at all times a matter of considerable
importance.
On two occasions I have noticed in lepers peculiar psoriasis-like
maculae, but in these instances the possibility of psoriasis and
leprosy occurring together could not be excluded.
REFERENCES
Abraham, Ph. (1910). Leproiy in * System of Medicine,* edited by Allbutt-Rolleston, Vol. II,
Part II, pp. 648-694.
Balfour, A. (1913). Leprosy in 4 Tropical Diseases Handbook, Ghent Exhibition,* pp. 90-104.
Castellani and Chalmers (1913). Leprosy in 4 Manual of Tropical Medicine,’ 2nd edition,
pp. 1147-1169.
Jadassohn, J. (1913). Lepra in 4 Handbuch der pat. Mikro-organismen,* pp. 791-930.
Manson (1911). Tropical Diseases, pp. 527-565.
III. MICROSCOPICAL FEATURES OF LEPROUS LESIONS
The minute anatomy of leprotic lesions or histo-pathology of
leprosy is the study, by means of the microscope, of the relationship
or direct and indirect action of the lepra-bacterium to and on the
tissues it has invaded. Accordingly, every investigation or analysis
of the appearances detected by the microscope must take into
consideration not only the presence or absence of acid-fast ‘ bacilli,’
but also, in the former eventuality, the quantity and localisation of
the ‘ bacilli ’ discernible in the sections being examined. Accordingly
it is to be understood that the microscopical diagnosis of a leprous
lesion is bacterioscopic and histo-analytic.
10
Good descriptions of the microscopical appearances of leprosy
have been given by Lie, MacLeod, Jadassohn and others, and an
even prolonged observation of the material available in South
Africa does not bring to light any new facts of fundamental
importance, except to show once more the necessity of clearly
distinguishing between leprosy and tuberculosis by means of
animal experiments and any other methods available for the
purpose. In referring to the lesions found in the spleens of lepers,
MacLeod (1910) expresses his view as follows (page 314):
'In the spleen, in addition to leprous infiltrations in the
trabeculae, pulp and malpighian corpuscles, extensive necrotic
masses have been described by Arning (Schaffer, Lepra, 1900, I,
p. n) like those of tuberculosis with typical Langhans giant cells.
Such cases have to be further verified from the point of view of
tuberculosis before they can be finally accepted/
In several autopsies (5) at Robben Island I have found in the
spleen of nodular lepers lesions similar to, if not identical with, those
described and illustrated by Schaffer. The inoculation experiments
into guinea-pigs were, however, positive; that is, the animals
experimented upon died within two to four months with extensive
generalized lesions of a tuberculous nature.
These investigations are by no means concluded, and they have
the limited value of single instances. They simply show that
‘ tuberculous 9 lesions in a leper are not necessarily always caused by
Hansen's ' bacillus 9 alone.
The uncompromising standpoint of Hansen, that leprosy and
tuberculosis were so absolutely distinct from each other that no
single common feature was thinkable, is no more defensible or
convenient, not even from a didactic point of view. At the present
day we have learnt through the more intimate knowledge of many
maladies and the properties of numerous pathogenic micro¬
organisms, that dissimilar diseases can be caused by microbes which
are practically indistinguishable, and that on the other hand
analogous pathological conditions can be brought about by micro¬
organisms which behave in a totally different fashion in artificial
culture and laboratory tests. As cases in point it is only necessary
to mention at random yaws and syphilis; kala-azar, infantile
splenomegaly and oriental sore; typhoid and paratyphoid; human
II
and bovine tuberculosis, plague and pseudo-tuberculosis. Pathology
has learnt the truth of Goethe’s dictum ‘ Alles ist Ubergang.’ The
increasing exactitude of clinical methods of observation ought to
enable us to avoid the pitfall of considering two allied diseases as
identical, since we understand that hard and fast types are only
extant for the purpose of classification.
For some time discussion has been rife whether leprotic tissue
ever contained giant cells of the Langhans type. It is now practi¬
cally unanimously admitted by those who have had opportunity of
seeing a sufficient number of slides from different cases, that giant
cells do occur in typical leprotic tissues, for they have been
observed by Thoma, Jadassohn, Ramon y Cajal, Rikli, Schaeffer,
Dohi, Babes, Kedrowsky, Matsuda, Gurd, Bayon and G. W.
Robertson (verbal communication). Even Lie, who originally held
that giant cells were only present in leprosy when the disease was
complicated by tuberculosis, now admits their presence, with the
reservation that they are not morphologically absolutely identical
to the Langhans type found in tuberculosis (verbal communication).
In discussing the nosology of leprosy, stress was laid on the
temporary nature of many conditions in leprosy, as a result Of the
extremely chronic course of the disease and the facility with which
it can be complicated by other infections. In addition we have to
consider the multifarious appearances of the disease per se. These
factors influence also the histological features to a considerable
extent, so that it is not possible to state, except in a somewhat
general fashion, which are the microscopical features that render
the diagnosis ‘ leprosy ’ certain in any section. No single element is
of decisive value, except the presence of numerous acid-fast rods,
and this even with several qualifications. Hansen’s ‘bacillus* is
generally only to be found in large numbers in advanced nodules
and nerve-lesions, and is practically absent from maculae and
leprotic papulae, that is, only to be detected after a very prolonged
search and by special methods.
Unna’s definition of a leproma is as follows: —
‘ .... a diffuse granuloma, whose peculiarity consists, on
the one hand, in its limitation to the connective tissue elements, and
specially to the lymphatic system of the skin, and on the other in
the enormous growth of organisms, whose number far exceeds
\ /
12
anything we are accustomed to find in other infectious diseases. . .
To the paucity of the cellular elements and the preponderance of
the organisms dormant in the bacillary mucus, the remarkable
indolence and relative benignancy of these growths may be
ascribed.* (Unna-Walker; The Histopathology of the Diseases of
the Skin, 1896, p. 616.)
Jadassohn sums up his investigations on the subject in the
following words* (l.c. p. 871): —
‘The leprosy bacillus produces a non-characteristic type of
inflammation, attended by the formation of granulomatous tissues
with different features of a degenerative character, which can even
deserve to be classified as necrosis; this may in some instances take
on a tuberculous appearance, signalised by sclerosis, and direct or
indirect destruction (Untergang) of various specific parenchymata.
As a rule the leprosy bacillus causes only an indolent reaction and
the resistance displayed by tissue against its luxuriant growth
(Wucherung) and toxines is quite remarkable. Deposits of bacilli
may be detected even in tissue which otherwise appears to be quite
normal, and in cells which cannot be shown to be altered in any
known fashion. The histological features are also very variable as
a result of the diversity of clinical types.’
‘ The typical leproma, wherever it may be situated in the body,
has the broad characteristics of other infective granulomata, with
certain peculiarities which render its diagnosis possible ’ (MacLeod).
It is evident from what has been said above that distinctive
features need not necessarily be present in many lesions which from
their occurrence and localisation we know to be definitely leprotic.
By avoiding generalisation, and considering single types and
anatomical situations by themselves, it is, however, possible to
recognise certain definite, recurring, important features.
Skin. 'Nodules. Numerous acid-fast rods occur, especially in
matted groups, there is absence of advanced necrosis (except in the
presence of open sores), and no caseation or giant cells with a
necrotic centre. Giant cells containing single acid-fast ‘ bacilli ’ may,
however, be present, but generally they are not numerous, and show
fewer peripheral nuclei than is the case in tuberculosis. The
‘ bacilli ’ may be scattered loosely in the parenchyma of the nodule
* Translated by the Author of this paper.
or may have invaded various cells of endothelial and fibroblastic
origin, in which case it is especially remarkable that the structure
of the protoplasm and nucleus or even the morphology of the
bacterial acid-fast rods has not suffered in any noticeable fashion.
Plasma cells are rare as opposed to yaws and syphilis, where they
are especially numerous.
A peculiar narrow band of hyaline, glassy tissue, containing no
‘ bacilli,’ is often noticeable just under the epidermis.
Acid-fast bacteria may be present in such quantities that in
stained sections (Ziehl-Nielsen) the whole tissue appears a dull red
to the naked eye. In other instances ‘ bacilli,’ though very
numerous, are distributed singly or by twos and threes, and can
only be detected with high-power lenses.
Skin. Patches. Acid-fast bacteria are not numerous. They
can only be detected with great difficulty. They are not bunched
or massed, but present in little groups of few ‘ bacilli,’ which may
apparently be situated extra-cellularly or they may have invaded
cells without causing any alteration in the structure of either
protoplasm or nucleus.
Giant cells with numerous peripheral nuclei arc often present.
At times a peculiar coagulation or discrete necrosis of the tissue
may be noted in parts, which shows hardly any definite structure.
It has a filamentary appearance and stains easily with eosin. Such
small areas of degeneration are circumscribed and show one or two
giant-cells at the periphery.
The vessels and capillaries can be seen partaking in a process of
proliferation, accompanied by an increase in the number of
endothelial cells forming their walls.
The epidermis is often found . thinner than usual, and the
epithelial villi are somewhat flattened out, or the epithelium may
appear as a border of uniform breadth.
Spindle-shaped cells (young connective tissue cells) may be
numerous, and plasma cells are not infrequent.
Skin. Maculae. Nodules and patches partake of the nature of
infectious granulomata. The macula belongs, however, to another
pathological group, which can be compared with a condition of
slight chronic irritation, possibly not very dissimilar to that seen in
Erythema perstans.
4
Various degrees of perivascular cellular deposit correspond to
the different intensity of eruption in single maculae, but, as a rule,
a slight infiltration of spindle and round cells can be detected
surrounding the capillaries and sweat-glands. The hair-follicles
may also undergo a similar process of localised infiltration and
proliferation. Giant cells may be present, but hardly any acid-fast
bacilli, unless nodules are to be found in other parts of the body.
The histological appearances I have described agree with the
view that maculae are a local anaphylactic symptom, caused by the
toxic products set free by the lepra-bacilli, after disintegration
induced through the agency of the defensive properties of the
tissues involved. This process takes place, however, when only few
germs are present, because lepra ‘ bacilli ’ in common with other
acid-fast micro-organisms are very resistant.
Regarding the changes occurring in the peripheral nervous
system, the suggestion has been made by Lie that ‘ bacilli ’ are only
present in maculae at the very beginning, and that later they
disappear into the nerve-endings and finally ascend along the
peripheral nerves. It can. however, be objected that in many cases
anaesthesia appears to set in without the primary production of
maculae in the corresponding area, and that many maculae do not
show any definite anaesthetic symptoms, though they have lasted
for years.
The other hypothesis considers the neuritis to be a descending
process (a contrast to the ‘ ascending ’ process put forward by Lie),
and accordingly the result of deposit following generalised
infection. In any case, though the nerves may be involved in any
single type of leprosy, it is in the maculo-anacsthetic type that this
invasion and localisation gives rise to the most striking and
apparently relatively rapid results.
In thickened peripheral nerves, under certain circumstances,
numerous ‘ bacilli ’ may be found, as in skin lesions. Generally
they are situated in massed groups in the spindle-cells of the
perineurium, or loose in the interstitial lymphatic spaces; at times
they may be distributed with a certain regularity in the cells of the
endoneurium, so that they appear to be in rows rather than in
clumps. In the course of time they may get eliminated and leave
behind a marked thickening, consisting of connective tissue with a
5
few stray degenerated and fragmented nerve fibres, in which the
general disposition of a nerve-bundle is still recognisable.
Practically every organ of the body has been found to be the
site of leprotic lesions by some observer or other, but deposits of
lepra-bacilli are more usually found in the lungs, liver, spleen and
testes. In the lungs the frequent complication with tuberculosis is
very disconcerting, and in many instances a decision as to the
pathological process involved can only be arrived at by tests with
inoculated guinea-pigs, though it is quite possible that even this
crucial experiment may not have an absolutely decisive value.
Babes gave considerable attention to the question of the lesions
found in the lungs of lepers, and has come to the following
conclusions which well deserve to be quoted in full :
(1) The lungs are normal, and do not contain lepra bacilli.
(2) They appear healthy to the naked eye and under the microscope, but
notwithstanding contain leprosy micro-organisms.
(3) They show the features of hypostatic pneumonia or broncho-pneumonia,
without any leprosy micro-organisms being recognisable, whilst
pneumococci and streptococci are present.
(4) They may show signs of tubercular localisation with excavations, caseous
pneumonia and peribronchial nodules, tubercle bacilli and without
the leprosy micro-organisms.
(5) In one single case he found caseous degeneration without any tubercle
or leprosy bacilli or other bacteria.
(6) In cases of chronic interstitial pneumonia peribronchial foci of a leprous
nature were noticed.
(7) The lungs may be the seat of extensive desquamative or fibrinous
localisation of a leprous nature, with countless lepra bacilli, which
may be accompanied by the formation of vomicae.
(8) Gangrenous bronchial cavities, and bronchial affections of a leprous
nature, were found in the centre of necrotic foci or surrounded by
an interstitial or desquamative pneumonia.
(9) Cases exist where leprous and tuberculous lesions combine, and
accordingly are very difficult to diagnose correctly.
The lesions of leprous lungs are very similar to those of tubercle, but it must
be remarked that leprosy runs a more chronic course, the resulting tissues are more
solid, caseate less frequently, and are less prone to be destroyed or eliminated.
When considered from this standpoint leprous lesions in the lungs resemble the
late manifestations of syphilis of the lungs.
We are thus enabled to distinguish not only a chronic diffuse interstitial or
trabecular form of leprosy of the lungs and a sclerotic nodular form, but also an
acute, and at times caseating, parenchymatous variety of the disease. In the
latter we also find fibrinous-pneumonic catarrhal lesions.
i6
In all cases he succeeded in proving, by means of cultures, and in some cases
also by microscopical examination in the bronchial forms, the presence of
diphtheroids in company with the lepra bacilli. These organisms are associated
at times and in varying manner with bacilli of the coli group ; at other times with
filamentary, branching micro-organisms or with streptococci or pneumococci in
the acute pneumonic or caseating varieties (‘ Lepra ; Handbuch der pathogenen
Micro-organismen ’ of Kolle-Wassermann, 1906, Erganzungsband, pp. 174-175).’
Leprotic lesions of the other organs may be accompanied by a
greater or lesser degree of amyloid degeneration. Amyloid of the
spleen, liver, kidneys, may be present in lepers who die with the
features of anaesthetic cases, but typical leprotic lesions of the
inner organs are generally only found in nodular cases.
Liver. The most frequent change noted is a peculiar interstitial
infiltration, accompanied by numerous acid-fast rods situated in
‘ lepra-cells,’ that is, mononucleated round cells whose protoplasm
is swollen out and riddled with vacuoles. The liver cells may also
contain ‘bacilli,’ though this is not frequent. Generally the
parenchymatous cells are but slightly altered, except for the
presence of lardaceous disease, and under certain circumstances may
contain a deposit of brownish-yellow pigment. The capillaries
appear dilated and may also contain lepra-micro-organisms, which
may have invaded the endothelium or lie loosely in the lumen.
In common with the other organs it is often very difficult to
decide in sections of the liver whether we are dealing with simple
deposits of ‘ bacilli ’ which have been carried by the blood stream
and have accumulated passively in the walls of the capillaries, or
whether we are dealing with a chronic but progressive pathological
process involving multiplication of the germs and an ever increasing
local spread of the leprous lesions.
In single cases giant cells and necrosis were detected, but I
fear that in these instances a complicating tuberculosis could not
be excluded. The same applies to the similar alterations seen in
the spleen.
Sfleen. In the spleen the acid-fast bacteria can be so numerous
that their presence can be detected by the naked eye on staining.
They may be scattered about in small clumps in the pulp,
especially in the neighbourhood of the capillaries. The presence
of lepra-cells may be under certain circumstances a noticeable
feature of the lesion. Though a spleen may be in an advanced
17
stage of leprotic invasion the resulting enlargement may be hardly
noticeable.
Testicles . The connective tissue between the tubuli is seen to
have proliferated actively, and to contain more or less numerous
acid-fast rods, some lying loosely, others enclosed in lepra-cells and
in the endothelium of capillaries.
Numerous degrees of connective tissue proliferation can be found
in the testicles from different patients, and under certain circum¬
stances even from different parts of the same testicle. The bacilli
present also vary very much in numbers—from few bacilli, which
can only be detected by means of an oil-immersion lens, to deposits
rivalling in numbers those seen in the nodules of some lepers.
Lymphatic glands . There is no doubt that the bronchial,
mesenteric, and other lymphatic glands can be affected by a pure
leprous infiltration, which microscopically shows very numerous
acid-fast rods enclosed in cells of different sizes, some with
hypertrophied protoplasm containing numerous vacuoles. Lymphatic
glands may also be the seat of a mixed infection with tuberculosis
and show necrosis and caseation.
From this brief survey it will be seen that in dealing with the
microscopical diagnosis of leprotic lesions we are continually faced
with the difficulty of distinguishing lepra from tuberculosis, and
that in many instances the decision must be left with the results of
inoculation into guinea-pigs.
It should not be forgotten that leprosy is a slower and more
chronic complaint than even tuberculosis, and that in certain
instances the possibility is given that a tubercle which has
spontaneously healed may be invaded by Hansen's 4 bacillus,’ in
which case the crucial experiment on animals would give a negative
result, though the histological lesions might show a definite
tuberculous character.
With the exception of cases where the presence of numerous
giant cells of the Langhans type, or very similar in appearance to
those seen in tuberculosis, coupled with necrosis or caseation,
render the presence of a mixed infection with tubercle probable,
the main microscopical features of a leprotic lesion can be summed
up in the following words:
Very numerous acid-fast micro-organisms, singly or in matted
i8
clumps, lie loose in the tissues and lymphatics or wedged in
compact masses in interstitial spaces. Similar acid-fast bacteria
may be present in varying numbers in the protoplasm of various
cells, in which under certain circumstances vacuoles can be detected.
The endothelial cells of the lymphatics may also show a marked
degree of bacterial invasion. The tissue reaction and production
of connective tissue is relatively insignificant in all instances, and
the resulting degeneration or necrosis of the tissues affected is quite
minimal in proportion to the number of germs present in the lesions.
REFERENCES
Aucanazy, M. (1911). Die Leprabazillcn. Lehrbuch der path. Anatomie herausgeg. von
Aschoff, I, 177-180.
Jadassohn, J. (1913). Lc.
Lit, H. P. (1911). Uber Tuberkuiose bei Leproesen. Archiv f. Dermat. u. Syphilis, CVII,
3-16.
MacLeod (1910). A brief survey on the present state of our knowledge of the bacteriology
and pathological anatomy of leprosy. Lepra XI, 309-320.
Schaeffer, J. (1900). Die Visceralerkrankung der Leprosen. Lepra I, 11-30 and II, 57-88.
IV. MORPHOLOGY AND STAINING PROPERTIES OF
HANSEN S ‘ BACILLUS 1
The Mycobacterium leprae belongs to a group of acid-fast
bacteria which in morphology and staining reactions are very
similar to the Mycobacterium tuberculosis . The group may be
considered to consist of the micro-organisms causing spontaneous
tuberculous disease in human beings, cattle, horses, pigs, birds,
etc. All these bacteria are relatively easily isolated on artificial
media, at body heat, from the lesions in warm-blooded animals,
with morphological and tinctorial properties analogous to those
which may be seen in infected tissues. A sub-division of this group
embraces several acid-fast germs, which are generally present in
diseased tissues in enormous quantities and are extremely difficult
to isolate in artificial culture. This sub-division may be considered
to include the Mycobacterium leprae , the M . leprae rodentium (rat
leprosy ‘bacillus’), and the micro-organism of Johne’s disease in
cattle, and that causing the analogous disease of sheep mentioned
by Twort and Ingram, Vukovic, McGowan. It is quite evident
*9
that a classification based on such principles cannot be satisfactory
from a bacteriological standpoint, but it is simply brought forward
to show once more that the leprosy ‘bacillus’ does not stand
absolutely alone in many of its peculiarities.
Lehmann and Neumann’s (1912) definition of the group
Mycobacterium is the following (p. 582):
' Thin, slender rods, often showing typical dichotomous
branching, at times giving rise to filaments, with or without
branching. If stained with warm carbol-fuchsine, the colouring
matter cannot be extracted by acids, that is to say, the bacteria
are acid-fast and behave towards stains somewhat like the spores
of common fission-fungi. In some species the acid-fast properties
are but slightly developed; in fact, may even be absent.’
According to Lehmann and Neumann’s definition, the myco¬
bacteria are not true * bacilli,’ and the possibility of micro-organisms
of the group showing true branching is recognised, as also the fact
that acid-fastness is a property which varies in different bacteria
belonging to the group. These admissions are important in view
of the fact that lately some bacteriologists have denied both
conclusions when applied to the microbe of leprosy.
A satisfactory classification of Hansen’s ‘ bacillus ’ is not a
matter of purely theoretical interest. It has definite practical
importance, because we have reason to expect that an exact
knowledge of its bionomics will help isolation by means of
artificial culture, and in such a fashion clear up the numerous
obscure points in connection with the epidemiology and therapeutics
of leprosy.
The irregular morphology of the micro-organism of leprosy
tends to confirm the view that it is not a true ‘ bacillus,’ because of
the lack of constancy in shape, thickness and length, which in
addition to various other features connected with the presence of
endogenous spores, flagella, etc., determines the diagnosis
‘ bacillus.’ The lepra bacterium is markedly pleomorphic, that is
to say, in smears from nodules single germs can be detected which
vary in thickness between o’3 ft and O '5 ft, and whose length may be
anything between 1 ft and 5 ft and in rarer instances even 6 ft or 7 /a.
Hansen’s ‘ bacillus ’ is often seen to contain small, fine granules
(Babes-Emst) which are not as acid-proof as the rest of the
20
bacterium. At times, peculiar hyaline breaks can be detected in
the rods; they have been considered to be spores.
Another peculiarity of the bacillus is its breaking up into small
coccoid bodies, which Deycke considered to be a sign of degenera¬
tion, but which the observations of Martinez-Santamaria seem to
indicate are possibly developmental stages prior to multiplication.
The reasons for which it is possible to classify Hansen’s
'bacillus’ as the acid-fast stage of a Mycobacterium closely allied
to the Actinomyces are based on morphological grounds and
deductions by analogy, in addition to the interpretation of
cultural attempts.
The morphology of Hansen’s ' bacillus,’ when examined with
high-power lenses (say 2 mm. apochromatic, comp, eyepiece No. 12)
and in sufficiently thin smears, can be seen to be exceptionally
variable; structures with one end tapering and the other definitely
thickened (clubbed) are mixed with rods of uniform thickness but of
different lengths and fragments, which seem to consist of a series
or chain of small beadlike coccoid bacteria.
At times in smears taken from nodules branching forms arc
seen; they are, however, rare. These peculiar pleomorphic
appearances, especially the clubbed and branching forms, are
typical of the fragments of actinomycotic germs having the
tendency to break up into acid-fast fragments. The variations in
morphology have lately been commented upon by Galli-Valerio,
who, in discussing the relationship of the genera Corynebacterium,
Mycobacterium and Actinomyces, has not failed to note the
numerous points of contact between these groups.
From the standpoint of analogy we know from the observations
of Petrone (1884), Metschnikoff (1888), Fischel (1892), Babes and
Levaditi (1897), Schulze (1899), Lubarsch (1899), Lehmann and
Neumann (1912), Galli-Valerio (1912), Foulerton, and others, that
true branching can occur in Mycobacterium tuberculosis and that its
appearances in sputum and tissues of experimental animals are at
times distinctly like those brought about by actinomycotic germs,
especially in the production of radiary deposits of bacteria. Abbot
and Gildersleeve have noted similar appearances in connection with
‘ saprophytic ’ micro-organisms. Further, observations of Silber-
schmidt (1899), Galli-Valerio (1910), Birt and Leishman, and
21
others, have made us acquainted with a group of germs which clearly
can be classed with the Actinomyces or Streptothrix class, yet in
artificial cultures and on injection into animals show numerous acid-
fast fragments very similar to M. tuberculosis.
That an acid-fast rod seen in tissues, can be found to be a
filamentary branching micro-organism in artificial cultures, is an
observation which has been made so repeatedly by those working
with A. caprae , Birt and Leishman’s Streptothrix, that there is no
necessity to dilate further on the subject.
Accordingly we may deduce that it is by no means far-fetched
to suggest that Hansen’s ‘bacillus’ may present a filamentary,
acid-labile appearance in artificial culture, though the M. tuber¬
culosis is consistently acid-fast on the usual laboratory media: and
even in this instance Frei and Pokschischewsky, and later Wherry,
have shown that under certain circumstances this property can be
made to vary.
The interpretation of the cultural results of investigators whose
bacteriological technique was not open to criticism (Beauchamp
Williams) shows us that in the great majority of cases their culture
tubes have either remained sterile for months and months (Fraser
and Fletcher), or that their repeated efforts were only rewarded by
the isolation of a singularly pleomorphic diphtheroid with slight
acid-resisting properties, that is to say, that after prolonged
staining with carbol-fuchsine it could be, and was, only partially
bleached by a weak acid such as one per cent, sulphuric. This
diphtheroid was usually found to have peculiar filamentary forms.
The relationship of the genus Corynebacteria to the Actinomyces
is also very close; to give an example out of a numerous series we
have the Corynebacterium ttecrophorum (Lehman and Neumann)
which Schmorl considers as a Streptothrix and other authors as an
Actinomyces.
As a matter of fact, the bacteriological literature of the last ten
years contains numerous, indeed very numerous, hints that a strict
separation of bacteria on cultural characteristics alone is extremely
difficult, to say the very least, because of the existence of very many
micro-organisms which do not conform in every case with any
single distinctive test or feature.
It will be seen that, though definite reasons are extant to
22
postulate that Hansen’s ‘ bacillus * belongs in reality to the Actino¬
myces, Actinobacteria or ray-fungus group, still the last conclusive
test is a matter of surmounting the difficulties inherent to the
artificial isolation and cultivation of this bacterium.
Staining Properties of Hansen’s ‘ Bacillus.’
In a recently published study on the bacteriology of human and
rat leprosy Wolbach and Honeij (1914) come to the conclusion that:
‘ The possibility that the non-acid-fast and acid-fast organisms isolated from
cases of leprosy are related and may be converted one into the other is one we
must take, however reluctantly, into consideration, because of the great length of
time the leprosy bacillus resides in the human body and the variety of conditions
it is subjected to by virtue of the stage of the lesion and anatomical situation.
The loss and acquisition of properties as distinctive as that of acid-resistance are
well known in the field of bacteriology and indeed the changes undergone by the
tubercle bacillus towards staining reactions is the strongest claim for the possibility.
Very numerous methods have been published with the purpose
of communicating a strictly specific stain for the leprosy micro¬
organism, that is, a stain which would show up in some distinctive
colour Hansen’s 4 bacillus,’ whilst all other bacteria, however similar
but not identical, would not take the same hue. The majority of
these methods are based on the property of the M. leprae of taking
up carbol-fuchsine or a similar stain more readily than Koch’s
1 bacillus,’ and, moreover, on the greater resistance, as a rule, of the
latter to the bleaching properties of mineral acids or strong alkalis.
Accordingly it will be seen that we are dealing with differences
in degree and not in quality, and as to be expected, such methods
of differentiation are apt to fail in a fair proportion of cases and be
unreliable in the remaining instances.
Our experience in dealing with the sections and smears in the
laboratory at Robben Island is that to obtain well stained leprosy
bacilli in sections it is advisable to use 2 \ per cent, carbolic in
making up the fuchsine solution, to stain in the warm solution one
or two hours according to thickness of section, and to differentiate
with one per cent, hydrochloric acid in the common methylated
spirit of commerce. Counterstain w'ith alum-haematoxylin.
Skin fragments are always fixed in absolute alcohol, specimens
from organs in 4 per cent, formaldehyde. Embedding in paraffin
is advisable, though excellent sections can also be got in celloidin.
23
For the quick staining of nasal smears, etc., Pappenheim’s
carbol-fuchsine-methylene-blue-coralline method gives reliable
results.
Unna has described a method of distinguishing lepra ‘bacilli’
which have been fixed alive from those that were brought dead into
the fluid. The validity of this distinction is, however, only founded
on circumstantial evidence.
REFERENCES
Foulerton, A. G. R. (1910). The Milroy Lectures on the Streptothrichoses and Tuberculosis.
Lancet, I, 551.
Frei und Pokschischewsky (1911). Centralbl. f. Bakt., LX, p. i6r.
Lehmann und Neumann (1912). Atlas und Grundriss der Bakteriologie.
Wherry (1913). Some chemical conditions influencing acid-proofness and non-acid-proofness
in a saprophytic culture of B. tuberculosis. Joum. of Inf. Dis., XIII, 144-154.
V. ARTIFICIAL CULTIVATION OF THE MICRO-ORGANISM
OF LEPROSY
The first attempts to cultivate Hansen’s “ bacillus ” were made by Sir Patrick
Manson many years ago. at a time when aniline dyes had not yet been introduced
into bacteriological technique. Later on. in 1882, Hansen thought he had
succeeded in getting the “ bacillus ” to grow artificially in some serum contained
between a microscope slide and slip, but he never followed the matter any further
after mentioning this observation.
Neisser in 1886 considered that multiplication of “bacilli” from a nodule
had taken place on cooked egg and blood serum. In any case he was not able to
get further generations to grow.
Bordoni-Uffreduzzi in 1887 cultivated from the bone marrow of a case of
leprosy a partially acid-fast or acid-resistant micro-organism, which, from the
description given and the specimens seen. I would consider to be morphologically
a diphtheroid. It was not quite as acid-fast as the Bacillus leprae in tissues.
Campana from 1889 onwards has described micro-organisms, which he
cultivated anaerobically from the nodules of lepers. I have been able to examine
the culture : it is a mixture of cocci, and coli-like organisms, none of which are
add-fast or even acid-resisting. There appears to be a sad lack of technique in
isolating these cultures, an opinion also apparently shared by Wasserman.
Kanthack and Barclay, in 1891, cultivated a micro-organism of varying add-
fast properties from a leproma. Originally they considered it to be leprosy,, but
later on they withdrew their claim.
Levy in 1898 cultivated from a leper a slightly add-fast, filamentary organism,
which in later publications he considered identical to that isolated by Czaplewski
in the same year from another case of leprosy. No animal experiments succeeded
with these micro-organisms.
Spronk. also in 1898, was successful in isolating a diphtheroid from lepromas.
He carried out some agglutination experiments with it, which, however, appear
2 4
not to have had conclusive results owing to incomplete knowledge of the
agglutinating properties of sera at that time.
Babes in 1899 isolated on several occasions, from nodules and the inner organs
of lepers, an acid-resisting diphtheroid, illustrations of which he has given, and
which I consider extremely suggestive. Unfortunately he did not follow the
work up, as he later considered that these diphtheroids were contaminations.
Barannikow in 1899 cultivated also a diphtheroid from lepers, very similar to
the one isolated by Bordoni-Uffreduzzi, Spronk, Babes, Levy, Czaplewsky. This
he considered was a pleomorphic stage of Hansen’s “ bacillus.”
Kedrowsky published his first results in 1900. Using placental-extract-agar,
he succeeded in isolating from the nodules of lepers an extremely pleomorphic
micro-organism, with varying acid-resisting properties.
Weil in 1906 succeeded in getting what he thought was a multiplication of
“ bacilli ” by injecting leper serum into a hen’s egg, which he incubated at 37°C.
He was not able to carry this further than one, the first, generation.
Rost published his first results in 1904. He made a great point of the fact
that the least trace of salt would not allow the culture to succeed. The medium
used was the extract from rotten fish. Semple in 1905 was sent to confirm Rost’s
results and found that his isolations were due to faulty bacteriological technique.
This was also the opinion of other bacteriologists who had occasion to examine
his work.
Rost’s culture is a dark red, wrinkled, dry culture. It is markedly chromogenic,
especially on gelatine at room temperature, under which conditions it grows quite
easily. At 37 0 C. it grows rapidly on most of the usual culture media, sometimes
within 48 hours.
Morphologically it is a slightly acid-resisting streptothrix, with add-fast
fragments ; this property varies, however, in different culture tubes. It is Gram¬
positive.
It resembles Grassbergeris isolation from butter in culture and under the
microscope. It has not produced leprotic lesions in animals.
Shiga (1909) isolated on potato-serum-glycerine a pleomorphic, partially
acid-resisting diphtheroid.
Clegg in 1909 published his results and concludes with the following summary :
1. The leprosy bacillus was first cultivated from leprous material in
symbiosis with other unidentified bacteria and amoeba, and later from
other cases in symbiosis with amoeba and cholera vibrion.
2. By heating a symbiotic culture of amoeba, cholera, and leprosy for
half an hour at 6o°C. and incubating, the leprosy barillus was obtained in
pure culture.
3. The leprosy bacillus, isolated in this manner, grows readily on the
ordinary laboratory culture media.
4. The bacillus is pathogenic for guinea-pigs, subcutaneous inoculations
having caused lesions, which macroscopically and microscopically resemble
the leprous lesions of the human subjects.
Clegg’s culture is granular, dry, yellow, grows at room temperature on gelatine
or any of the common laboratory media. I have found difficulty in getting the
strain to grow in the incubator. It is add-fast, alcohol-fast, and Gram-positive.
Morphologically it consists of short plump rods, very similar in appearance to the
timothy grass bacillus ; at times, however, it may resemble the ‘ 4 Smegma bacillus,”
and accordingly consist of slender matted rods.
25
On injection into animals it does not produce lesions comparable with leprosy
in man.
Currie, Brinckerhoff and Hollmann in 1910 succeeded after several failures,
in growing acid-fast rods in symbiosis with cholera vibriones and amoebae. They
divided up their cultures into four “ strains,” and found that one was practically
identical with Clegg’s culture from Manila.
Serra (1910) cultivated anaerobically from several cases (three of seven
attempted) a peculiar micro-organism which is Gram-positive, but not acid-fast,
as it can be bleached by 2 per cent, nitric acid. Morphologically it appears to
be a “ diphtheroid.”
Twort in 1910 by using his “ ericoline ” method isolated from the nasal
discharge of a leper, on egg-medium to which he had added ground-up tubercle
bacilli, an acid-fast rod, which grew very slowly indeed, and was morphologically
identical with the “ bacilli ” found in tissues. He was not able to carry the culture
further than the first generation, but still his incomplete results deserve considera¬
tion, in view of his success with Johne’s “ bacillus.”
Duval in 1910 and the following years published various results in connection
with the isolation of Hansen’s “ bacillus.”
His first publication was founded on Clegg’s results, and the isolation of the
acid-fast micro-organisms was made with amoebae in the fashion identical with that
of Clegg. The micro-organism in question appeared, however, to differ from
that isolated by the latter. In later publications Duval found he could dispense
with the amoebae and cultivate the “ bacilli ” from the nasal discharge of lepers
simply by adding amino-acids to the culture media. Later on, even the addition
of the amino-acids does not seem to be indispensable, for an isolation is mentioned
on Novy-Mac Neal’s blood-agar. The last papers deal with isolation by means
of Kedrowsky’s placental juice agar, which had been re-discovered by Wellman
and published as “ Wellman’s placental agar.” Animal experiments failed to
produce typical leprous lesions.
Duval’s so-called culture of lepra is a bright yellow, moist, smooth culture.
It grows easily and abundantly on gelatine by room temperature and can be easily
regained in pure culture on common agar from the organs of animals that have
been inoculated with it. At 37 0 C. it takes about three days to grow on any of
the current laboratory media.
Morphologically it is coccoid in appearance, but under circumstances, when
injected into animals and sometimes on artificial media, it is apt to present a
somewhat more elongated appearance, but at no time closely resembles Hansen’s
“ bacillus.”
It is acid-fast and alcohol-fast. Gram-positive.
Its cultural and morphological characteristics cause it to resemble markedly
a * saprophytic acid-fast micro-organism isolated by Nabarro from London milk.
Therefore it is not astonishing that animal experiments did not succeed in
producing any lesions analogous to those found in lepers, and that at the
International Medical Congress in London (1913) this author admitted he had
no culture of leprosy to show or hand round.
Kedrowsky’s paper published in 1910 deserves special mention in view of
the care with which the experiments have been carried out. They took ten years
to complete.
His first isolations from the nodules of three lepers gave as a result two distinct
bacteria; one was a non-acid-fast filamentary, interlacing, branching micro¬
organism, the other was a slightly acid-resisting diphtheroid. He also cultivated
26
other micro-organisms, which, however, from his pictures and descriptions are
easily seen to be but variations of these two groups. He injected these various
bacteria strains singly, into mice and rabbits, in different fashions (under the dura,
intravenously and intraperitoneally) and observed the animals for prolonged
periods, in some cases over two years. He found that, whatever micro-organism
he had injected, the result was practically the same in all cases; very numerous
acid-fast micro-organisms of the “ tubercle bacillus ” type in the viscera, from
which they could be regained in pure culture as acid-fast rods. The resulting
lesions resembled in some cases the chronic type of tuberculosis induced in rabbits
by injections of human tuberculosis ; in others the lesions were strikingly similar
to those occurring in visceral leprosy of human beings.
Kedrowsky’s acid-fast culture of “Hansen’s Bacillus” is a moist, creamy,
smooth or wrinkled ivory-white culture which resembles avian tuberculosis to a
very marked extent. It grows only at incubator temperature on special media,
such as placental-juice-agar, glycerine-agar, horse-serum-nutrose-agar or any
similar medium suitable for tubercle. Multiplication is generally apparent within
ten to fourteen days, but may take three weeks to four weeks or more to attain
its maximum.
No growth takes place at room temperature or on gelatine.
It is add-fast, cannot be bleached by 20 per cent, nitric add in one minute
after staining for five minutes with warm carbol-fuchsine. It is alcohol-fast, will
withstand absolute alcohol for ten minutes. Gram-positive. Morphologically it
is markedly pleomorphic, and resembles avian tuberculosis in this respect, in fact
I do not believe that these two bacteria can be distinguished from one another
under the microscope, except that Kedrowsky’s strain of lepra is more apt to show
clubbed shapes than even avian tubercle.
Williams in 1911 isolated diphtheroids and streptothrices from the nodules
of lepers. He considered them to be pleomorphic stages of Hansen’s “ bacillus,”
but was not able to produce any typical lesions in animals with the micro-organisms
isolated.
Currie, Clegg and Hollmann express their opinion on these cultures in the
following terms:—
“ This investigator considers that his work tends to confirm the work of Rost,
Deycke, and Clegg, i.e., that all these investigators have, by different methods,
isolated the same organism, which latter is very pleomorphic. He seems, however,
to base this opinion on very little data other than that the organisms in question
were all grown from lepromata, and that local nodules were produced in animals
by inoculation of some of these cultures.”
Bayon (1911) published the first confirmatory results of Kedrowsky’s isolations,
above all in relation to the existence of filamentary non-acid-fast micro-organisms
in cultures taken with all aseptic precautions from leprous nodules. He also
confirmed the possibility of isolating on various media diphtheroids which were
slightly acid resisting. Both bacteria acquired acid-fast properties after injection
into animals. This author considered that the identification of bacteria isolated
from lepers was the most important point in connection with the bacteriological
work, and accordingly carried out extensive investigations by means of numerous
experiments on animals, serological tests and observations on human beings. He
found that Kedrowsky’s view that the parent form of Hansen’s “ bacillus ” is a
filamentary, interlacing, branching, non-acid-fast micro-organism is correct, and
that this pleomorphic property is shared by several other acid-fast bacteria, e.g. ;
the smegma “ bacillus,” some saprophytes, such as the acid-fast rod found in
tap-water, etc. His comparative experiments with Duval’s. Rost’s, Clegg’s cultures
27
persuaded him that they behave in culture, on common gelatine at room tempera¬
ture, and in animal experiments, like the usual ubiquitous, acid-fast micro-organisms
found in grass, dung, milk, butter, earth, etc. This observation he confirmed by
serological tests, which showed that these cultures when used as antigens do not
react specifically with any lepers’ serum. The lesions produced in animals are
not like the lesions of rat or human leprosy. On the other hand, he found that,
given sufficient time for incubation, and notwithstanding a very heavy percentage
of failures, Kedrowsky’s strain was capable of producing in rats, mice and rabbits
lesions extremely similar to those occurring in visceral lepra of human beings. To
obviate as much as possible any experimental error, the culture was injected into
guinea-pigs and fowls to determine whether, as Babes had suggested, it might not
be a tubercle strain which had infected Kedrowsky’s animals. Small doses (five
to ten loopfuls) did not cause any lesions in guinea-pigs or fowls, even after pro¬
longed observation.
The serological tests employed also showed this strain to be capable of reacting
specifically with the serum of certain lepers. A filtered cultural extract when
injected into lepers caused a rise of temperature to take place. This reaction,
however, has no specific value, as it can be brought about by many different sub¬
stances, and is, moreover, not constant.
Bayon’s culture (in the diphtheroid stage) was studied by Priestley, who com¬
pared it with numerous (namely, 48) other micro-organisms from various sources,
showing somewhat similar morphological features, with the intention of bringing
evidence to bear on the point, whether this diphtheroid could possibly be an
ubiquitous saprophyte, whose presence in the skin of lepers had only casuistic,
and not etiological importance.
Priestley found that morphologically the “ bacilli ” were very irregular in
shape, clubbing was very marked, polar granules could be detected by Neisser’s
stain. Gram-positive. On serum they formed raised, white circular colonies,
like Klebs-Loeffier “ bacillus ” ; on agar, very minute, circular, greyish-white
colonies; no growth on gelatine; on broth, very slight growth, very stringy
deposit; no growth on potato. Non-virulent. They fermented dextrose and
laevulose, but not saccharose, lactose, maltose, mannite, dulcite, glycerine, dextrite,
galactose, did not clot milk. Unfortunately the study was not completed at the
time, and no further details were published because all attention had to be con¬
centrated in attempting to bring the peculiar results of some American authors
in relation with the observations made on Kedrowsky’s culture and the acid-fast
micro-organism isolated from rats inoculated with acid-labile cultures from lepers.
Numerous experiments on animals showed, however, that the acid-fast stage
of the filamentary micro-organism was capable of producing in rabbits, rats and
mice lesions very similar to those obtained by Kedrowsky in his experiments, and
which accordingly were analogous to the lesions observed in leprosy of the human
being.
The result of these numerous experiments, prolonged over several years (4),
was accordingly that Kedrowsky’s isolation was true leprosy, and that the cultures
of some of the other authors appear to be accidental.
Reenstierna (1913) succeeded in cultivating from the blood and nodule of a
leper an acid-fast rod, which was morphologically similar to Hansen’s “ bacillus ”
seen in tissues, and numerous acid-labile bacteria. The acid-fast rods were
isolated in pure culture by the means of 10 per cent, antiformin, and lasted four
generations in their add-fast condition, and then turned acid-labile.
A pure culture isolated from these acid-labile bacteria showed on single
occasions the presence of acid-fast micro-organisms.
28
A monkey (Macacus rhesus) was injected in the brain and ischiatic nerve with
a pure culture of the acid-fast rod isolated from the blood of a leper. This animal
showed peculiar maculae on the chest and face, which contained acid-fast micro¬
organisms. The death of the monkey took place three months after injection,
and the inner organs showed numerous caseating nodules which contained big
bundles of acid-fast rods, which were mostly situated extra-cellularly.
Another Macacus rhesus was injected with a pure culture of the acid-labile
micro-organism isolated from the blood of a leper. Forty-two days after, it
developed blisters on its fingers and toes, which contained acid-labile and acid-fast
bacteria. Shortly after the animal was killed, and acid-fast rods were found at
the site of injection, and in caseating inguinal and sacral lymphatic glands.
A third M. rhesus was injected with a culture of the acid-labile micro-organism,
which was isolated from a single bacterium by means of Bum’s Indian ink method.
This animal lived eight months, and though during its life it developed a spot
which contained diphtheroids and a contracture, the autopsy revealed no leprotic
lesions.
Reenstierna concludes from his numerous observations that the bacillus of
leprosy is not only morphologically but also biologically closely related to the
tubercle “ bacillus.”
His experiments deserve special consideration, because they belong to the few
which have been undertaken without the preconceived idSe fixe that leprosy and
tubercle are absolutely distinct diseases in every single particular, and that Hansen’s
“ bacillus ” is acid-fast from beginning to end of its existence, notwithstanding
that there is sufficient evidence to show that either statement need go unchallenged.
Stanziale (1913) cultivated from the eye of a rabbit, which had been inoculated
128 days before with a fragment of leprotic nodule under anaerobic conditions, on
agar-egg-yolk, an acid-fast rod very similar to that of tuberculosis. The original
culture took a month to develop, but on other media the colonies were already
apparent after twenty-four to forty-eight hours. On agar and broth the culture
resembles tubercle, except for the fact that it is more abundant. Apparently the
resistance to the bleaching properties of acids varies, though the bacterium contains
granules which are markedly acid-fast. Gram-positive. Numerous clubbed
shapes similar to those seen in Corynehacterium diphtheriae. Non-pathogenic for
animals.
Wolbach and Honeij (1914) have lately isolated a diphtheroid from a case of
leprosy. It was originally cultivated aerobically on ascitic fluid dextrose agar,
and appeared within ten days in the form of a translucent whitish band around
the piece of gland tissue.
“ When first isolated the growth was poor on bouillon, with or without glycerin,
and on plain agar, dextrose agar and glycerin agar. After several months’ cultiva¬
tion the bacillus grew readily on these media. No growth has been obtained
upon potato, with or without the addition of 0*5 to 1 per cent, sodium carbonate.
Glycerin does not favour the growth of this bacillus.” Gram-positive. After
staining with carbol-fuchsine (how long ?) they resisted decolourization with 1 per
cent, hydrochloric acid in 70 per cent, alcohol; 20 per cent, sulphuric acid left the
bacilli from litmus milk distinctly red, after staining by Gabbett’s method for
tubercle. No lesions were obtained on inoculation into Japanese waltzing mice,
guinea-pigs, rabbits and white rats. The methods of inoculation were intra¬
venous, subcutaneous and intraperitoneal.
Wolbach and Honeij consider this culture to be identical with the diphtheroid
which has so often been isolated from cases of leprosy, and put in a plea for more
full data about any micro-organisms which may be cultivated in future.
2 9
These are the results obtained in the course of upwards of thirty
years of bacteriological investigation of an extremely widely spread
disease. No doubt hundreds of negative results have been obtained
and not published, but it can be said that as a rule every time the
problem has been tackled by a competent bacteriologist the culture
and isolation of a diphtheroid or filamentary germ has been the
result. This has also apparently been the case with the investiga¬
tions of Fraser and Fletcher, who, however, discarded all
diphtheroids because of their ubiquity, a standpoint which would
considerably simplify pathology. The micro-organisms of syphilis,
of tuberculosis, of cholera, could also, on similar grounds, simply
be ruled out of court, as far as their etiological significance is
concerned.
The main point has, however, always been that as a rule
these diphtheroids, though slightly or partially acid-resisting,
did not turn absolutely acid-fast on injection into animals, with the
exception of the instances mentioned.
It should be understood that every experiment connected with
leprosy should be repeated scores of times, because evidently if the
matter were simple it would have been solved from the very
beginning. The crucial experiment to be carried out with any
culture isolated from a leper is not only to get it to acquire the
acid-fast properties and the morphology of the ‘ bacillus * seen in
tissues, but also to succeed in producing lesions analogous to those
seen in lepetS and in rats spontaneously infected with M. leprae
tnurts. Bearing in mind the great difficulties encountered in
transmitting leprosy to animals through the injection or inoculation
of nodules teeming with ‘ bacilli/ we must be prepared to face series
and series of negative results. As negative results, however
numerous, are powerless to destroy a single positive observation, it
must be admitted that Kedrowsky has published micro-photographs
and drawings of lesions in rabbits and mice which are strikingly
like those produced by the inoculation of human and rat * virus 1 in
rabbits and rats. Such specimens have not been published by any
other author, excepting those to be found at the end of this paper;
the corresponding deduction should not be difficult. (Plate II,
fig. no
3 °
REFERENCES
(Arranged in chronological order.)
Nusser, A. (1886). Histologische und bakterioiogische Lepra-unter*uchungen. Virchow’*
Archiv, CIII, 355.
Bordoni-Uffriduzzi, G. (1887). Uber die Kultur der Leprabazillen. Zeit*chrift fur Hygiene,
III, 178.
Manson, Sir P. (1890). Journal of the Leprosy Investigation Committee, I, 40.
Stallard, J. H. (1890). The Lepra Bacillus. Occidental Medical Times, IV, 220.
Campana, R. (1891). Un badllo simile al badllo leproso sviluppatosi in tentativi di cultura di
tessuti con lepra tuberculare. Riforma medica, No. 14.
Ducrey, A. (1892). Tentativi di cultura del badllo della lepra con risultato positive. Giornale
italiano delle malattie veneree, etc.
Levy, E. (1898). Ein neues, aus cinem FaBe von Lepra gezfichtetes Bakterium a us der Klasse
der Tuberkelbadllen. Archiv ffir Hygiene, XXX, 168.
CzAPLEWsrr, E. (1898). Uber einen neuen, aus einem Leprafalle gezfichteten, alkohol- und
sSurefesten Bacillus aus der Tuberkelbadllengruppe. Centralblatt ffir Bacterio logic,
XXm, 97 and 98.
Spronk, C. H. H. (1898). De cultur van de badl van Hansen en de serodiagnostik van lepra.
Nederl. Tijdschr. van Geneeskunde, II, 322.
Teich, M. (1899). Beitrage zur Kultur des Leprabadllus. Centralblatt fur Bakteriologie,
XXV, 756.
Babes, V. (18^9). Uber die Kultur der von mir gefundenen Diphtherideen. Centralblatt fur
Baktenologie, XXV, 12$.
Barannikow, J. (1899). Zur Frage uber die Bakteriologie der Lcpromata (Vorlaufige
Mittheilung). Centralblatt ffir Bakteriologie, XXV, 113.
Kedrowsky, W. J. (1901). Beobachtungen fiber die kfinstliche Kultur des Lepraerrcgers.
Zeitschrift ffir Hygiene, XXXVII, $2.
-(1904)* Experiraentelle Erfahrungen fiber Lepraimpfungen bei Tieren. Centralbl.
ffir Bakteriologie, XXV, 368.
- (1910). Experimentelle Untersuchungen fiber Lepraimpf ungen bei Tieren.
Zeitschrift fur Hygiene, LVT, 1.
Van Houtum, G. (1902). A successful attempt to cultivate Bacillus leprae. Journal of
Pathology, VIII, 260.
Zenoni, C. (1902). Ricerche batteriologiche sulla lebbra. Gazzetta medica italiana, LIII,
No. 45 -
Levy, F. (1902). La coltura artifidale del bacillo della lepra. Giornale della Sodeta italiana
d’lgicne, XXIV, 219.
Gjubert, J. (1903). Zur Bakteriologie der Lepra. Baumgarten’s Jahresbericht, XIX, 337.
Karlinsky (1903). Kulturen eines aus Lcprabaciilen gezuchteten saurefesten Bacillus.
Monatshefte ffir praktische Dermatologie, XXXVII, No. 9.
Rost, E. R. (1905). The cultivation of the Bacillus leprae. Indian Medical Gazette, February.
Deycke und Reschad (1905). Neue Gesichtspunkte in der Leprafrage. Deutsche mcd.
Wochenschrift, No. 13, p. 489. No. 14, p. $45.
Weil, E. (1906). Essais de culture du bacillus lepreux. Annales de l’lnstitut Pasteur, 1905,
793 -
Clegg, M. T. (1909). Some experiments on the cultivation of Bacillus leprae. Philippine
Journal of Sdence, IV, 77.
-(1909). The cultivation of the leprosy bacillus. Ibidem, IV, 403.
Currie, Brinckerhoff* and Hollman (1910). On the cultivation of the bacillus of leprosy
by the method of Clegg. Public Health Reports, page 1173.
3i
Duval, C. W. (1910). The cultivation of the leprosy bacillus and the experimental production
of leprosy in the Japanese dancing mouse. Journal of Experimental Medicine, XII, 649.
- (1911). The cultivation of the leprosy bacillus from the human tissues, with special
reference to the amino acids as culture media. Journal of Experimental Medicine,
XIII, 365.
Twort, F. (1910). A method of isolating and growing the lepra bacilli of man. Proceedings
of the Royal Society, LXXXIII, 156.
Skr&a, A. (1910). Contributo alio studio del bacillo di Hansen. Giora. ital. Mai. ven. Fasc. II.
-(1911)* Inoculation de culture du bacilli de Hansen dans l’oeil du lapin. Lepra,
XII, 1-14.
Williams, T. S. B. (1911). Leprosy: A new view of its bacteriology and treatment.
Supplement to the Indian Medical Gazette, May, p. 1.
Bayon, H. (1911). Demonstrations of specimens relating to the culture of the leprosy bacillus.
British Medical Journal, November nth.
- (1912). A comparative experimental study of the leprosy cultures of Clegg, Duval,
Kedrowsky, Rost and Williams. British Medical Journal, November 2nd.
- (1914). The Micro-organism of leprosy, has it been cultivated ? Lepra, XIV, 187-192.
Liston, W. G. and Williams, T. S. B. (1912). A Streptothrix isolated from the spleen of a
leper. No. 51, Scientific memoirs (Med. and Sanitary Depts.) Govt, of India.
Rzenstieina, J. (1913). Uber die Kultivierbarkeit und Morphologic des Lepra-erregers und
die Ubertragung der Lepra auf affen. Archiv fur Dermatologie, CXVI, 1-76.
Stanzialx, R. (1913). Isolamento colturale di un bacillo acido resistente da occhio leproso di
coniglio. Atti della R. Accademia Napoli, No. 2, pp. 3-7.
Wolbach and Honuj (1914). The diphtheroid bacillus from leprosy lesions. Journal of Med.
Research, XXX, pp. 1-8.
VI. THE EXPERIMENTAL TRANSMISSION OF LEPROSY
TO ANIMALS
Leprosy is definitely known to be a bacterial disease, and
therefore it should not and cannot be impossible to transmit it to
animals by experimental inoculation, if it is considered that practi¬
cally all pathogenic bacteria can be made to produce in animals
diseases similar or analogous to those caused in man. In fact, the
existence of a disease in rats, extremely similar to leprosy, ought
to throw light on this vexed question and postulate the possibility
of the experimental infection of laboratory animals.
In estimating the value of any experiments made for the purpose
of inoculating animals with leprosy, it should be remembered that
but fourteen years ago it was universally denied that syphilis could
be given to rabbits or monkeys. Even scientists like A. Neisser
upheld this belief, which, as we now know’, was absolutely
erroneous and totally unfounded. Syphilis is extremely contagious,
develops comparatively rapidly, therefore it is not astonishing that
32
the transmission to animals should be a relatively easy matter, much
more so than leprosy. Yet, even after the communication of the
first successful inoculations of syphilis to rabbits and monkeys,
numerous negative experiments were published, accompanied by
the statement that the sores produced on the testis of rabbits, etc.,
were not produced by the syphilitic virus but were due to the effects
of contaminating bacteria.
These recent happenings should induce more attention being paid
to the evidence which has accumulated in the course of time,
showing that though in the great majority of cases leprotic
products, when inoculated into animals, are eliminated without
leaving any definite traces, still multiplication and conclusive
dissemination of the bacilli does take place in a small number of
instances.
In 1881, Neisser, after twenty-four unsuccessful attempts on rabbits,
injected two dogs with leprous nodules, and concluded from the results that leprosy
developed locally at the site of inoculation.
1883. Damsch grafted lepromata in the anterior eye chamber of two rabbits.
The animals died after 139 and 219 days. Apparently a slight increase in the
quantity of acid-fast germs had taken place. He also inoculated two cats in the
abdomen with leprous nodules. After 120 days numerous bacilli were found in
the newly-formed tissue round the atrophied nodules.
1888. Vossius carried out experiments in a similar fashion to Damsch on
rabbits and had identical results. He also considered that the acid-fast germs had
multiplied.
1885-6. Melcher and Ortmann inoculated four rabbits in the same way.
They succeeded in getting, after four to ten months, definite metastases and deposits
of acid-fast germs in the spleen, liver, caecum, pleura, and pericardium. They
stained these germs with Baumgarten’s differential stain, and found that they had
the staining properties of Hansen’s “ bacillus.”
1887. Wesener injected several rabbits with leprous nodules, and in two
animals out of eight, after periods of four and a half months and eight months
respectively, he found lesions apparently identical with those described by Melcher
and Ortmann—that is, nodules from the size of a pinhead to that of a pea in the
lungs, on the pleura, on the epiploon, in the liver, in the lymphatic glands, spleen,
kidneys, caecum, and peritoneal surface. Caseation and giant cells were present.
In the eyes of the rabbits he obtained after six and eight months a congregation of
round cells choked up with acid-fast micro-organisms. He considered that the
generalised lesions were due to tuberculosis, and that the ocular lesions were caused
by dead bacilli, because acid-fast germs can be demonstrated in the eye of the
rabbit after inoculation with nodules which have been kept for years in alcohol.
1901. Barannikow confirmed these results on one rabbit.
1893. Wnoukow inoculated twenty rabbits with leprous nodules in various
fashions (intraocularly, subcutaneously, intraperitoneally). In fourteen rabbits
he got lesions which he considered to be of tuberculous nature. No confirmatory
tests with guinea-pigs appear to have been made.
33
1893. Tedeschi inoculated a leproma into the dura of a monkey. The
animal died after eight days; that is far too brief a period for any conclusions to
be drawn.
1902. Ivanow injected several guinea-pigs with leprous nodules. In one
case the animal was killed after eight months and found to have nodules in the
epiploon, which Ivanow considered were due to multiplication of the bacilli.
1905. Thiroux inoculated five rabbits with leprous material. The animals
lived thirteen to twenty months, and presented various lesions at autopsy, which
Thiroux considered to be tuberculous, after positive inoculation of two guinea-
pigs.
1906. Nicolle inoculated monkeys with ground-up nodules, and succeeded
in getting localised lesions.
1909. Marchoux and Bourret inoculated a chimpanzee under the skin of
the ear with a freshly-excised leprous nodule. It apparently increased in size
during three months, and then began to be resorbed. An examination of the
blood showed a few leucocytes with badly-staining bacilli. The animal died ninety-
six days after inoculation. The nodule had reached the size of a split pea. and under
the microscope was shown to consist of three layers. In the middle were the
remains of the original tissue, quite necrosed, with a few loose acid-fast germs.
Around this was a considerable number of lymphocytes, enclosing numerous bacilli,
and at the periphery organised connective tissue with a few cells filled with bacilli.
The authors considered that a proliferation of the original germs had taken place.
1909. Sugai inoculated Japanese dancing mice intraperitoneally with an
emulsion of fresh leprous nodules, and found as a result the development of miliary
granulomata on the peritoneal lining, especially in the hepatic region and epiploon.
Also the bronchial and peritoneal glands showed characteristic leprous lesions. All
contained acid-fast germs. The attempt to transmit the disease from these to
other mice failed.
1909. Kitasato appears to have successfully inoculated an orang-outang on
the cornea. Only scanty details were, however, given.
1909. Stanziale reports upon experiments on rabbits. In one typical case
he inoculated into the eye of the animal a fragment of leproma. During the first
few days a marked hyperaemia developed round the opening in the cornea. Soon
afterwards a constant diminution in the size of the nodule could be made out,
accompanied by the formation of an opalescent exudation, which took its departure
from the inoculated piece of tissue and spread out as a fan-shaped segment. After
twenty days the nodule became stationary, and remained so during fifteen to
twenty days without any noticeable change taking place. After forty days the
nodule began slowly to increase till it had trebled its original size. In the same time
small grey nodules made their appearance, and could clearly be distinguished on the
surface of the inoculated tissue. These little nodules attained the size of a pin’s
head. Some appeared to be adherent, while others were loose and independent.
After the initial hyperaemia due to the operation had disappeared, a new formation
of blood vessels took place, which spun a delicate network round the piece of tissue
inoculated. After seventy days the rabbit’s eye was enucleated, part of the tissue
was used for histological observations and the rest was grafted into the cornea of
two other rabbits. In one the graft got resorbed without any further results in
the course of two months. In the other rabbit the graft began to increase in size
after fifty days, and produced similar changes to those described in the foregoing
animal. The experiments are being continued by the author, but to date the
results obtained are : thirty-one rabbits have been inoculated, and in eight it was
possible to produce leprous ocular lesions. In some of the positive cases the
3 +
material was taken eighteen to nineteen hours after excision from the patient. In
all cases in which the inoculation succeeded the Wassermann reaction was positive.
As a control Stanziale inoculated lepra nodules into the abdomen of another rabbit,
and found at intervals after a month the reaction constantly negative. The same
negative result was achieved after grafting into the cornea of rabbits pieces of human
slrin or dead leprous nodules.
These experiments of Stanziale have been most carefully carried out, and full
descriptive details are given ; the only defect is, however, that no metastases in the
inner organs are mentioned.
1911. Puval inoculated a series of animals—namely, four rats, four white
mice, and four Japanese dancing mice—with grumous material taken from an
acute case of human leprosy which had developed numerous soft subcutaneous
leprous masses and had repeated attacks of leprous fever. A small quantity of
material was used in each case—namely 0*5 c.cm. emulsified in 1 c.cm. of normal
saline solution. Some were injected intraperitoneally, others subcutaneously.
The two white mice which received intraperitoneal injections died fourteen
days after the inoculation. At the autopsy both showed a general infection of the
peritoneum with a pure growth of the M. lepra, while the mesentery, omentum,
visceral and parietal peritoneum contained numerous minute, firm greyish-white
nodules, which on microscopic section proved to be typical leprous lesions. The
most surprising feature was the occurrence of a slightly turbid, semigelatinous
exudate, which microscopically consisted almost entirely of large mononucleated
cells (macrophages). Great numbers of these cells were filled with acid-fast bacilli,
and scarcely any were found that did not contain a few. No remarks are made
about the other animals.
1911. Couret fed two goldfish on teased nodules from a leper. No other
food was allowed the animals until they had disposed of the leprosy tissue, after
which they were cleansed and transferred to a clean aquarium.
The first was examined twenty-four days later; acid-fast organisms had not
been found in the faeces for three days before the animal was killed. No evidence
of infection was apparent externally or internally. Films made from the omentum
and other organs showed a few scattered acid-fast bacilli. The average number of
bacilli to the slide was about four, and no change in morphology was seen. The
second fish was examined after thirty-seven days. The findings were similar to
those in the first one. With reference to these experiments, it should not be
forgotten that tap water appears to harbour acid-fast germs frequently, as shown
by Brehm, Beitzke, Schern, and Dold. The tap water at the Lister Institute in
London and of the Public Health Laboratory in Cape Town, also contains acid-fast
germs; I have also seen them in the taps at Stockholm. (See Plate IV, fig. 20.)
Serra (1911 and 1913) describes a series of experiments he has carried out on
rabbits by means of the intra-ocular method.
His results on the whole seem to correspond to those of Stanziale, but it is
extremely difficult to unravel any definite statements as to the facts observed
from a mass of secondary detail. Too much stress seems to be laid on the Wassermann
reaction observed in some of the rabbits inoculated, and no details are given as to
way in which it has been carried out; whilst also in this case no metastases in the
inner organs are mentioned.
It is to be hoped that other following publications will bring more light on
the subject.
Ch. Nicolle and Blaizot succeeded in producing in Macacus sinieus , by inocu¬
lating fragments of leprous tissue, local lesions very similar to those of dermal
35
leprosy in man. They repeated these experiments on three monkeys (bonnet
chinois) and one chimpanzee, and reported on the results in 1911.
One monkey received sixteen inoculations in toto> but none of them ever
generalised, and the resulting nodules got slowly resorbed in the course of time.
A second monkey died without showing any visceral lesions. The experi¬
mental lepromas were in course of development, and showed numerous acid-fast
micro-organisms under the microscope.
A third monkey received nine inoculations at intervals varying between a week
and twenty-five days. The resulting nodules softened, caseated and broke down.
The pus showed a few degenerated “ bacilli ” among numerous polynuclear cells.
The chimpanzee received eight inoculations in the course of four months.
One injection was followed by the development of a local nodule, which was
followed later on by several little nodosities, which soon became confluent. This
lesion only lasted about a fortnight and then got resorbed.
In these cases, therefore, we had a temporary or negative result in most cases,
and a localised success in one experiment.
Bayon (1912) injected four rats on two occasions in the testes with ground-up
nodules from a case of leprosy from Mauritius. Two rats did not show any macro¬
scopic lesions, even after four months. One rat, however, developed, four weeks
afterwards, a nodule at the site of inoculation, which grew to the size of a small pea.
On puncture it showed acid-fast germs and necrosed tissue, some of which, no
doubt, represented the original cells injected. After five weeks the rat died, but
no acid-fast micro-organisms were found in the organs. Microscopical sections
were made of liver, spleen, and testes. The disease, if transmitted, was localised.
The whole of the nodule was then injected subcutaneously into two other young
rats. Three months after inoculation, one of the rats was losing fur on the surface
of its abdomen, without there being any signs of the usual rat scabies about the
ears or tail. Small shotty nodules could be felt under the skin.
The other rat developed, three months after the second injection, a nodule
in the left testis. This broke down, and was found to contain acid-fast rods in
great quantity. This rat was killed four months after inoculation, the slight
wound caused by the nodule breaking down having healed. At the site of injection
and in the corresponding inguinal glands numerous acid-fast rods were found.
One of these small glands was inoculated into the testes of another rat, and on
killing it six months afterwards no acid-fast micro-organisms were found in the
testes, but definite deposits were present in the inguinal glands, spleen and liver.
A multiplication must have taken place, because the acid-fast bacteria detected in
these organs were evidently more numerous that the relatively small quantity
injected. (Plate III, Fig. 14.)
At Robben Island, during 1912, 1913 and 1914, thirty experiments on rabbits
were undertaken by means of the intra-ocular method. Exact data will be
published in due course, when the histological study of the resulting specimens is
undertaken.
It is, however, worthy of note that one single rabbit of the thirty inoculated
has shown permanent lesions—that is, minute white, vascularised nodules in
the cornea and on the iris. This animal was operated upon 1st November, 1912 ;
nodule from Pat. No. 1294.
As a rule the remaining animals developed inflammatory appearances similar
to those described by Stanziale, but in the course of time the inoculated fragment
of leprotic tissue got resorbed, all signs of inflammation or production of new
tissue subsided, and in conclusion only a linear scar remained as a trace of the
operation.
3 6
One animal, however, still shows numerous minute leprotic lesions one year
and seven months after inoculation.
I do not wish to admit a definite communication of leprosy to these animals,
till I have been able to achieve the results accomplished with the rats—that is,
the transmission of the virus through several generations, and deposits in the inner
organs.
Reenstierna (1912) injected a Macacus rhesus under the mucous membrane of
the nose with 0*5 c.cm. of an emulsion made from a leproma containing numerous
“ bacilli.” The animal died forty-two days after inoculation. The inner organs
showed no alteration, but the nose had developed a perforation of the septum ;
moreover, there was extensive ulceration of the upper lip and nasal mucous mem¬
brane. The microscope revealed numerous extra-cellularly situated acid-fast rods
in the nasal secretion and sections of the diseased tissues. In addition an acid-
labile but Gram-positive filamentary germ was present with fragments which,
morphologically, were similar to lepra “ bacilli.”
Another M. rhesus was injected intra-peritoneally and subcutaneously with
the same emulsion as in the previous experiment. The monkey died fifty-nine
days afterwards, and showed numerous caseating nodules in the liver, spleen,
lung and mediastinal lymph-glands. These lesions contained numerous acid-fast
rods.
Five guinea-pigs were inoculated with the caseating nodules from the spleen ;
they died within three to five weeks and showed localised lesions which contained
very numerous acid-fast rods.
A third M. rhesus monkey was injected with the same emulsion intraperi-
toneally and subcutaneously. It died six months afterwards, and developed small
nodules at the sites of injection which contained acid-fast rods. A cherry-sized
nodule which developed on the upper-lip contained numerous acid-fast intra¬
cellular “ bacilli ” and vacuolated cells. Acid-fasts were also found in the liver,
spleen and bone-marrow.
These are the successful or reputed successful attempts to
transmit leprosy to animals by means of injection or inoculation of
leprous tissues.
As will be seen, they are certainly more numerous and have been
more carefully observed, than the isolated statements in connection
with the communication of syphilis to animals, which were to be
found in literature only so short a time ago as the first year of this
century. And yet syphilis can be communicated to rabbits with
relative ease. In dealing with leprosy, where even direct inocula¬
tion of human beings has failed (except in Aming's case) one can
but expect to find much more serious difficulties, such as:
(1) Lack of knowledge in relation to the bionomics of Hansen's
‘ bacillus.'
It does not seem to be understood that though attendants in
leper asylums, and persons living with lepers, must often come in
contact with numerous ‘ bacilli ’ which we know nodular lepers cast
37
about them, yet only a very small proportion ever develop the
disease in a clinically recognisable form. In other words, in the
great majority of cases either the ‘ bacilli ’ remain absolutely
quiescent in the body or get harmlessly eliminated.
Therefore it should not astonish us that a great proportion of
the animals inoculated eliminate the acid-fast bacteria injected after
a longer or shorter period.
(2) The difficulty of proving whether any ‘ bacilli ’ seen in
experimental lesions are simple deposits of dead acid-fast bacteria
or whether they are alive and in course of multiplication.
This is really an outcome of No. i, as it depends on an
incapacity in comprehending how extremely passive, inert and
resistant Hansen’s ‘ bacillus ’ is.
It is true that Campana, Wesener and others have shown that
it is possible to bring about lesions in animals very similar to those
described by the experimenters who claim having transmitted
leprosy to rabbits or rats, by simply injecting tissues containing
numerous ‘ bacilli ’ which had been autoclaved or kept in alcohol
for months. As we know of no method of cultivating directly the
acid-fast germs seen in leprosy with their original morphology and
staining properties, it is of course difficult to refute this argument
except by pointing to the analogy with rat-leprosy, where, though
we know that the ‘ virus ’ used in experiments is living because it
•can be indefinitely transmitted, still, except for the latter particular,
the lesions caused by killed and living virus are extremely similar.
(3) The similarity existing between the lesions produced in
rabbits and those found in experimental tuberculosis.
This is the argument which settled the results of Melcher and
Ortmann, Wesener and others. At the time leprosy and tuber¬
culosis were considered to be as easily distinguishable histologically
and bacterioscopically as they are clinically. Further study has,
however, shown that this is not the case, and that under certain
conditions only experiments with guinea-pigs can decide whether
we are dealing with lesions brought about by Koch’s or Hansen’s
' bacillus.’
Taking into careful consideration the previous experiments and
personal experience, in addition to the interpretation of the clinical
notes made on lepers who have been under observation for fifteen
3 «
and more years in South Africa/ no other conclusion can be arrived
at than that leprosy is directly, experimentally, communicable
from human beings to rats and rabbits; that a long time of
incubation must be allowed for; that the great majority of animals
eliminate the 1 bacilli * injected after a shorter or longer period of
quiescent deposit.
We have, therefore, under experimental conditions an analogous
counterpart of what is happening daily with people who contract
leprosy, for even prolonged exposure to infection causes the disease
to develop only in a small percentage. Those who show clinical
symptoms of the contagion appear to eliminate the majority or
practically all their 1 bacilli * if they live long enough to become
what is called an 4 arrested * case. Such an 1 arrested * case is a
leper who is known to have had numerous visible lesions teeming
with acid-fast rods, but who for the time being only shows scars
and traces of the disease with no 4 bacilli.* Such cases relapse at
times, or may show small bacillary deposits in their inner organs
at death, but even then there can be no doubt that they have
succeeded in eliminating or destroying millions and millions of
4 bacilli * from or in their nodules.
A somewhat analogous observation is often made at autopsies
in connection with tuberculosis of the lungs. It is then seen that
very many bacillary infections of this organ do not develop into
clinical tubercles, but heal with scarring.
All these considerations should, and do, give a satisfactory
explanation of many negative results and condone the scepticism
with which many successful experiments have been received.
Conclusive histological specimens like those shown in Plate III,
fig. 14 cannot be ignored, especially as experimental results agree
in an exceptional manner with clinical observation.
Once more, these conclusions have to be arrived at :
(1) Leprosy is experimentally transmissible to animals such as
the rabbit and the rat. Allowance must be made for the fact that
the great majority of inoculations do not succeed, owing to the
4 bacilli * being eliminated or destroyed without leaving any visible
trace.
• This shows that in single instances lepers get rid of nearly all the acid-fast germs from their
lesions.
39
(2) The incubation period is very long, and the resulting
lesions in the inner organs may not be visible to the naked eye.
(3) In rabbits the bacillary deposits in the inner organs
resemble tuberculosis. In such cases the question can only be
decided by means of guinea-pig inoculations.
REFERENCES
(Chronologically arranged)
Neisser (1881). VVeiterc Bcitrage zur Aetiologie der Lepra. Virchow’s Archiv, LXXXIV, 314.
Damsch (1883). Ubcrtragungsversuche von Lepra auf Thiere. Virchow’s Archiv, XCII.
Melcher und Ortmann (1885). Ubertragung von Lepra auf Kaninchen. Berliner klinische
Wochenschrift, p. 293, No. 13. (1886) Experimentelle Darm und Lymphdriisen Lepra
bei Kaninchen. ibidem No. 9.
Wesenek (1888). Zur Frage der Lepraiibertragung auf Tiere. Centralblatt fiir Bakteriologie,
III, 482.
Vossius (1888). Zeitschrift fiir vergleichende Augenheilkunde, VI, 1.
Wnoukow (1893). Inaug. Thesis Kasan (Russian), Ref. Centralblatt fur Bakteriologie,
XII, 173*
Tedeschi (1893). Centralblatt fiir Bakteriologie, XIV, 113.
Iwanow (1902). Sur lc sort des bacilles de la lepre dans l’organisme des animaux (cobayes).
Annales de l’lnstitut Pasteur, No. 10.
Y’ amada, Toyama, Kurita (1903). Japanese Journal of Dermatology, III, 529, quoted by
Sugai.
Thiroux (1905). Quelqucs tentatives d’inoculation de la lipre. Annals d’hygiene et mid.
coloniale, VIII, 148.
Nicolle (1906). Recherches experimentales sur la lipre. Annales de l’lnstitut Pasteur XX,
389.
Marchoux et BoURRtT (1909). Recherches sur la transmission de la lipre. Annales de
l’lnstitut Pasteur, XXXIII, 513.
Sugai (1909). Nachtrag zur gelungenen Ubertragungsversuchen mit Lepra bei Saugetieren.
Lepra, VIII, 203.
Kitasato (1909). Die Lepra in Japan. Zeitschrift fiir Hygiene, LXIII, 507.
Stan zi ale (1910). Inoculazioni di materiale leproso nella camera anteriore dei conigli.
Giornale italiano delle malattie veneree, V, 1.
Couret (1911). The behaviour of bacillus leprae in cold blooded animals. Journal of
experimental medicine, VIII, 576.
Serra (1911). Inoculation de culture du bacille de Hansen dans oeil du lapin. Lepra, XII, 1.
Duval and Gurd (1911). Studies on the Biology of and Immunity against the Bacillus of
leprosy. Archives of Int. Medicine, VII, 231.
Nicolle, Ch. et Blaizot (1911). Essai de reproduction de la lipre chez le Chimpanz6 et les
Singes inferieurs. Archives de l'lnst. Pasteur de Tunis, 275.
Bayov (1912)- On the transmission of leprosy to animals by direct inoculation. British
Medical Journal, February 24th.
Nakano (1912). Uber die kunstlichc Zuchtung von Leprabazillcn in Tierleichen. Archiv.
fur Dermatologie, CXI, 819.
- (1912a). Experimentelle Untersuchungen iiber die Infektionsmoglichkeit von
Japanischen Hausratten, Kaninchen und Meerschweinchen mit Lepramaterial. Ibidem,
CXIII, 787.
4 °
On Rat Leprosy and its Relation to the Human Disease
A disease extremely similar to leprosy, not only in its clinical
features but also from an histological and bacteriological point of
view, was discovered in rats by Stephansky in Odessa, in 1903. Its
presence among the sewer rats of Berlin was confirmed by Lydia
Rabinowitch the same year. Both these observers considered that
rat leprosy could not be transmitted experimentally from one
animal to the other, and that the acid-fast micro-organism found
in enormous quantities in the lesions of the animals affected could
not be cultivated on nutrient media.
George Dean had studied this same disease at Elstree, near
London, for some time, and published his results nearly con¬
temporaneously with Stephansky and Rabinowitch. He was,
however, successful in transmitting rat leprosy through several
generations of rodents, a result which has since been amply
confirmed by Marchoux and others. G. Dean’s cultural attempts
only succeeded in isolating a diphtheroid, which showed filamentary
forms, and much to his astonishment reacted specifically with the
serum of lepers. The results of his agglutination experiments
were :
1 1. The serum of rats inoculated with rat-leprosy, i.e., with acid-fast
“ bacilli,” agglutinated the diphtheroid. Normal rat’s serum had no agglutinating
properties for this micro-organism.
2. The serum from a case of human leprosy agglutinated the acid-fast micro¬
organism from the rat. Normal human serum had no agglutinating power.
3. Normal human serum and the serum from a tuberculous patient failed
to agglutinate the diphtheroid, whereas the serum from a case of leprosy had
distinct agglutinating properties.’
These carefully planned tests, in addition to the histological
appearances, the cultural and experimental behaviour of the micro¬
organism causing the disease in rats, lead us to believe that rat
leprosy is possibly related to human leprosy in a manner similar to
that existing between bovine and human tuberculosis.
Marchoux has carried out very carefully-planned experiments
with the strain of rat leprosy he discovered in Paris, and has come
to the following conclusions: —
* 1. That only six in a thousand sewer rats in Paris present marked leprotic
lesions, but that 5 per cent, harbour acid-fast micro-organisms (bacille de
Stephansky).
2. That the inguinal glands appear to be the first attacked by the disease.
4 1
3. That the lung acts as a filter in arresting the germs and then directing
them towards the mediastinal glands.
4. That by simply placing the virus of the disease on the scarified or depilated
skin, an infection can be brought about in rats even more easily than by subcutaneous
injections.
5. That intact skin or even the bare skin of very young rats does not allow
the penetration of the virus.
6. Rat leprosy is a disease peculiar to the rat. Mice can be infected, it is
true, but with less ease than is the case with rats. Attempts to infect through
the scarified or depilated skin do not succeed as often as subcutaneous injection in
mice. In such cases one finds amongst the “ bacilli ” involution forms similar to
those met with in rats injected with human leprosy.
7. Mycobacterium leprae murium , like Hansen’s “ bacillus ” is a parasite of
the mesodermal cells. It does not live at the expense of its host, but of the same
substances which nourish the cell containing it.
8. Granular bacilli are dead.
9. A first culture is relatively easy to obtain. Further sub-plants are not
obtainable.
10. The bacillus disappears rapidly in contaminated media (en milieu impur).
11. It does not resist desiccation.
12. It is not killed by a temperature of 6o°C. during five minutes. It dies
after being exposed at the same temperature for fifteen minutes.
13. Infection takes place through the skin, and accordingly superficial surface
regions are generally more intensely affected.
14. Infection follows the lymphatic stream.
15. The point of inoculation is not always the most heavily infected spot.
16. Male rats can be infected by depositing some of the virus under the
prepuce without causing any lesions of the mucous membranes.
17. However, the spontaneous disease does not seem to be transmitted in
this fashion.
18. Insects do not carry the disease.
19. The sarcoptes of rat scabies play only an indirect role in the diffusion
of the disease.
20. The immediate contact of a skin abrasion with a diseased surface or
with freshly contaminated objects is the usual mode of contagion.
21. By introducing a large number of micro-organisms through the digestive
tract a primary pulmonary infection can be produced.
22. The lymphatic glands of artificially inoculated rats are generally very
small; those of spontaneously diseased rats, on the contrary, generally much
enlarged.
23. Inoculation generally produces an infection of the lymphatic glands.
Impure virus has to be injected to produce the muscular and cutaneous type of
the disease.’
I have quoted extensively tiiese results of Marchoux and Sofel,
because not only all experiments have been carefully and
systematically worked out, but the majority of the observations
made seem to agree so completely with what we know of the
pathology of human leprosy.
4 2
Currie and Hollmann also carried out experiments with rat
leprosy, and in two papers on the subject came to the following
conclusions:
* i. In some cases of artificially acquired rat leprosy the onset is with broncho¬
pneumonia, accompanied by a septicaemia and without any other demonstrable
lesions.
2. In other cases of this disease pneumonia is a very early lesion, but we
cannot positively state that it is always the first lesion.
3. That the animal may die in the pneumonic stage before other lesions
present themselves, or it may develop pneumonic symptoms and recover from the
same only to develop later the well-known lesions.
4. That during the stage of the disease in which the animal is very ill certain
mites (Latlafs echidninus) were found to be very numerous on the animals’ bodies.
5. That during the stage of the disease in which septicaemia is marked, these
mites* digestive tracts contain the bacilli of rat leprosy in considerable numbers,
and, therefore, these parasites may be one means of transmitting the disease. This
latter probability is, of course, not proven.’
In a later paper, published in 1912, these authors add the
following to the previous conclusions:
‘ 1. In the disease we are dealing with, whether the animal is inoculated by
a laboratory method or simply allowed to develop the disease from coming into
contact with infected rats, i.e., the natural mode of infection—the lesions met with
are practically the same.
2. With the exception of the local lesion, occasionally produced at the site
of artificial inoculation, infection of the viscera seems to usually precede the lesions
of the skin.
3. Of the visceral lesions, a broncho-pneumonia is often the earliest and
most constant. Infection of the spleen is also often an early event.
4. The heart blood of infected rats often contains the bacilli of rat leprosy,
and no difficulty is experienced in demonstrating the presence of acid-fast bacilli
in the mites contained on the bodies of these animals when the latter’s heart blood
contains the organisms.
5. The fact that these mites contain the bacilli so frequently naturally leads
one to suspect that they may be one of the means of transmitting the disease from
rat to rat, but up to the present time we have no evidence that such is the case.
Attempts to Cultivate the Bacillus of Rat Leprosy
As reported in our first paper, we have made many attempts to cultivate the
bacillus of rat leprosy on artificial culture media, both on the media ordinarily
employed in the laboratory and by the method of Clegg in symbiosis with amoeba
and cholera. We have now to report that we have continued these experiments
for‘nearly a year, and that our results have been entirely negative, not having
secured a single culture of an organism which we considered to be the bacillus of
rat leprosy. In one of our attempts to cultivate this organism from the ulcer on
the abdomen of a leprous rat, we succeeded in growing what appeared to be an
acid-fast streptothrix. As one might expect to obtain such organism on an
exposed surface of that kind, we regarded this culture as an accidental contamina¬
tion, and not the bacillus of rat leprosy.’
43
It is hardly feasible to bring these results in correlation with
anything observed in human leprosy, except for the fact that also
these authors have found the disease can be transmitted by means
of inoculation from one animal to another, an experiment which
did not succeed in the hands of earlier observers. In a similar
fashion the transmission of human leprosy to animals has failed
time after time, and yet under exceptionally favourable circum¬
stances it can be successfully achieved. The repeated attempts to
cultivate the micro-organism of the disease, which only resulted in
the isolation of acid-fast filamentary, branching bacteria (diph¬
theroids are not mentioned) which were considered to be
contaminations, are also worth noting, because this observation
coincides with the results of much of the work done on the
bacteriology of human leprosy.
Bay on (1912) succeeded in isolating on fish-juice-agar from the
spleen of a rat which had been injected with a ground-up nodule
from a spontaneously diseased rodent, a moist, ivory-white, creamy
growth, which multiplied in sub-cultures very slowly at 37 0 C. on
the media generally used for the artificial cultivation of tubercle.
This culture produced in rats the identical glandular and visceral
lesions observed in rats injected with the virus from spontaneously
diseased animals. The glands and organs infected were teeming
with acid-fast micro-organisms, which caused little or no tissue
reaction and a very minimal production of giant cells (Plate III,
figs. 16 and 17).
The culture does not grow at room temperature or on common
gelatine.' It is difficult to isolate again from the inner organs of
the animals into which it has been injected. Guinea-pigs, fowls and
rabbits do not show any lesions if inoculated with small quantities
(five to ten loops), but if a culture slope or more is introduced intra-
abdominally, after a time caseating, necrosed nodules are produced
on the surface of the peritoneum and the omental glands can be
considerably enlarged, showing big masses of acid-fast micro¬
organisms. The necrosis and caseation are partially due, I believe,
to the mechanical foreign-body action of the large numbers of
resistant micro-organisms injected.
No growth could be obtained under anaerobic conditions.
In an oxygen atmosphere the cultures develop somewhat more
44
rapidly; after ten to fourteen days a thick growth is discernible,
whilst under ordinary conditions, at 37 0 C., three or four weeks are
needed.
No multiplication was obtained on gelatine at room temperature
or on common agar in the incubator.
No pigment was produced on any of the following media:
fish-j uice-glycerine-agar, glycerine-agar, glucose-agar, glycerine-
potato, rice-glycerine-milk, sterilised wedges of rabbit’s liver +
glycerine, Dorset’s egg-medium. All these media were suitable for
growing the micro-organism.
Morphologically the culture is a short rod, with a marked
tendency to pleomorphism; clubbed shapes are frequently seen, and
at times true branching can be detected. The length of the rods
varies; in fact, the microscopical appearances are very similar to
those of a culture of avian tuberculosis, which it also resembles
from a cultural point of view.
It is acid-fast, i.e., cannot be bleached by one minute’s treatment
with 20 per cent. H 2 S 0 4 after having been stained for five minutes
with warm carbol-fuchsine. It is also alcohol-fast. After soaking
for six hours in warm alcohol its staining properties are unimpaired.
It is Gram positive.
Very numerous attempts to isolate the same micro-organism from
several other cases of rat leprosy failed completely. Marchoux is
therefore not quite correct in saying that: ‘ Bayon, au contraire,
l’aurait cultiv6 assez facilement.’ (l.c. p. 23.)
Further cultural results have been published by Hollmann,
Chapin, Zinsser and Carey, Marchoux and Sorel, and, of course,
Wellman. None of these cultures have succeeded in producing
leprotic lesions.
After Stephansky’s and Rabinowitch’s and Dean’s observations,
rat lepra was found in Australia by J. R. Bull, in New South Wales
by Tidswell, in the United States by Wherry, McCoy, Walker, in
Roumania by Mezinescu and Alexandrescu, in Japan by Jitoyo and
Sakaki, in Paris by Marchoux, in New Caledonia by Leboeuf, in
Ipswich by Petrie and Macalister (see PI. I, fig. 4), in India by the
Plague Commission, in Queensland by Priestley.
Though rat lepra seems to be spread nearly all over the world,
still Brinckerhoff examined 16,000 rats in the Hawaii Islands
without finding any trace of an animal affected with the disease.
45
In Cape Town I have had now 1,378 sewer rats examined
without having succeeded in a single instance in discovering a
marked case of the disease.*
Dr. G. W. Robertson remembers, however, to have noticed one
animal the lesions of which contained numerous acid-fast rods, in
the course of plague investigations, when about 30,000 rats were
examined.
Mezinescu succeeded in preparing from the ground-up nodules
of leprous rats an antigen which was capable of absorbing comple¬
ment when linked up with the serum of lepers; a further proof of
the relationship of both diseases, though, on the other hand, it
must be admitted that the serum of lepers is similar to that of
syphilitics in this respect, because it is capable of reacting with
specific and certain non-specific antigens. A distinction can only
be made by carefully titrating all the elements of the haemolytic
system, and varying the proportions of antigen and antibody in a
suitable fashion; up to the present this has only been done in quite
a minimal proportion of cases.
Rats are not the only animals affected by a disease akin to
leprosy. Acid-fast micro-organisms which belong to the tubercle
group, but are not pathogenic for guinea-pigs in small quantities,
and are only cultivable with extreme difficulty, cause diseases in
cattle, horses, sheep, and possibly goats. The cattle disease has
been named by K. F. Meyer 4 Enteritis hypertrophica bovis
specifica.*
Twort and Ingram, Vukovic, MTadyean, McGowan, have
described 1 Johne’s disease* in the sheep. This disease differs from
rat-leprosy in so far that the symptoms are mostly confined to the
digestive tract.
REFERENCES
(Chronologically arranged.)
Stephansky, W. K. (1903). Eine lepraahnliche Erkrankung der Haut und dcr Lymphdriisen
bei Wanderratten. Centralblatt fur Bakteriologie, XXXIII, 481.
Rabinowitch, L. (1903). Uber eine durch saure-feite Bakterien hervorgerufene Haut-
erkrankung der Ratten. Centralblatt fiir Bakteriologie, XXXIII, 577.
Dean, G. (1903). A disease of the rat caused by an acid-fast Bacillus. Centralblatt fiir
Bakteriologie, XXXIV, 222.
- (1905). Further observations on a leprosy-like disease of the rat. Journal of Hygiene,
v, 99- _
*My thanks are due to Dr. Mitchell and Dr. J. Anderson of Capetown for procuring
these rats.
4 6
Tidswell, F. (1904)* Note on a leprosy-like disease in rats. Report of the Board of Health
on Leprosy in New South Wales, p. 13.
Reports on Plague Investigation, etc. (1907). Journal of Hygiene VII, 337.
Bull, J. R. (1907). Leprosy-like disease of the rat. Intercolonial Med. Journal of Australasia,
May 20th, p. 215.
Wherry, W. B. (1908). The leprosy-like disease among rats on the Pacific Coast. Journal of
the American Medical Association, L, 1903.
- (1908a). Notes on rat leprosy and on the fate of human and rat lepra bacilli in flies.
Journal of Infectious Diseases, V, 307.
- (1909). Experiments on vaccination against rat leprosy. Ibidem, VI, 630.
McCoy, G. W. (1908). Leprosy-like disease in rats, U.S.P.H. and Marine Hospital Service
Reports, July 10th.
- (1908a). Distribution of the leprosy-like disease of rats in San Francisco. Ibidem,
November 6th.
- ( I 9 I 3 )* Observations on naturally acquired rat-leprosy. Ibidem, No. 61, XXIV,
27-30.
Mezinescu, D. (1908). Maladie 16 preuse des rats et ses relations avec la lipre humaine.
Comptes rendus de la Soci6t6 de Biologie, LXIV, 314.
- (1909). Ibidem, LXVI, 56.
Walker, A. (1918). A report on some cases of rat leprosy. Journal of the American Medical
Association, LI, 1153.
Alexandrescu, J. (1908). Lepra sobolanilor. Quoted from Marchoux (1912).
Kitasato (1909). Die lepra in Japan. Zeitschrift fur Hygiene, LIII, 507.
Brinckerhoff, W. R. (1910). Rat leprosy. U.S. Public Health Bulletin, No. 30.
Currie, D. H. and Hollmann, H. T. (1910). A contribution to the study of rat leprosy.
U. S. Public Health Bulletin, No. 41.
- (1912). XVII. Further observation on rat leprosy. Ibidem, No. 50.
Jitoyo, K., and Sakaki, R. (1910). On rat leprosy. Ref. Centralblatt fur Bakteriologie,
LVIII, 690.
Ehlers, Bo urret et With (i 91 i). Recherches sur le mode de propagation et les proc£des de
diagnostique bact6riologique de le lipre. Bulletin dc la Soci6t6 de Pathologie Exotique.
3 *» 2 53 -
Leboeuf, A. (1912). Existence de lepra murium en Nouvelle Caledonie. Bulletin de la
Soci6t6 de Pathologie Exotique, No. 7.
Marchoux, E., et Sorel, F. (1912). La Wpre des rats. (Lepra murium.) Annales de
l’lnstitut Pasteur, Aout.
-(1912a). Lepra murium. Comptes rendus de la Society de Biologie, LXXII,
159, 214, 269.
Marchoux, E. (1912). Role des infections secondaires dans le developpement de la lipre du
rat. Bulletin de la Society de Pathologie Exotique, p. 466.
- (1912a). Human leprosy and rat leprosy ; a discussion of their respective problems.
Trans. Soc. of Trop. Medicine, V, 184.
Bayon, H. (1912). The culture and identification of the germ of leprosy and the relationship
of the human disease to rat leprosy. Transactions of the Society of Tropical Medicine,
V, 158-167.
-(1912a). A comparative experimental study of the cultures of Clegg, Duval, •
Kedrowsky, Rost and Williams. British Medical Journal, Nov. 2nd.
Meyer, K. F. (1912). Bacillus para-tuberculosis Johne. Proceedings Philadelphia Pathol.
Society, October.
M’Gowan (1914). A case of Johne’s disease in the sheep, in ‘Monograph on Scrapie,* page
113-U4.
Wolbach and Honeij (1914). A critical review of the bacteriology of human and rat-leprosy.
Joum. of Med. Research XXIV, pp. 367-423.
+7
VII. THE SEROLOGY OF LEPROSY AS A MEANS OF DIAGNOSIS
OF THE DISEASE AND FOR THE PURPOSE OF CONTROLLING
SPECIFIC TREATMENT
Soon after the adaptation of Bordet-Gengou’s complement
fixation to the diagnosis of syphilis by Wassermann and his
co-workers, Eitner modified this test to suit the special conditions
of leprosy, and prepared his antigen by means of a saline extract
from a leproma. He found that the blood of lepers contained, as
he thought at the time, specific anti-bodies, and accordingly was
able to bring about the absorption of complement when linked up
with an antigen prepared according to his method. In the course
of time it became evident that Wassermann’s test for syphilis could
not claim to be absolutely specific* Accordingly Eitner amplified
his experiments, and found that the serum of lepers was also
capable of giving a positive reaction when linked up with the
alcoholic extract from a guinea-pig’s heart, and that therefore, also
in this particular instance, the serum of lepers and that of
syphilitics behaved in a similar fashion.
Numerous and various experiments were then carried out with
different antigens by several investigators, and the serum of lepers
was carefully compared with that of syphilitics and tubercular
patients. As far as the original Wassermann test of syphilis is
concerned, its relation to leprosy can be aptly summed up in the
conclusions of Howard Fox (l.c.): —
‘ I. A positive Wassermann reaction is frequently obtained in cases of
leprosy giving no history or symptom whatever of syphilis.
2. The reaction is at times very strong, inhibition of haemolysis being com¬
plete.
3. The reaction occurs chiefly in the tubercular and mixed forms of the
disease, especially in advanced and active cases.
4. In the cases of the maculo-anaesthetic and purely trophic type the reaction
is generally negative.
5. The value of the test is not affected in the slightest by the results found
in leprosy.’
On the other hand, if we consider the test being made with an
antigen prepared in various fashions from the nodules of lepers
(Eitner’s test or its modifications), one finds on reviewing the
literature on the subject that, with a few differences, the results
mainly agree with Jeanselme’s conclusions, which are the following:
48
‘ i. Eitner’s reaction applied to syphilitic sera is positive in 85 per cent, of
all cases examined.
2. Eitner’s reaction with the serum of healthy individuals or that of syphilitics
in whom the disease is quiescent: 100 per cent, negative.
3. Wassermann’s reaction applied to lepers: Practically the same conclusions
as Howard Fox.’
Eliasberg has shown that the serum of lepers is at times capable
of inhibiting haemolysis by itself, even small quantities as 0*4 c.c.
being sufficient. A similar observation has been made by Ehlers
and Bourret.
Noguchi’s 4 Luetin * reaction was found to be negative in eleven
lepers who were free from obvious signs of syphilis (but gave a
Wassermann reaction) by Clegg.
Incidentally it can here be mentioned that, according to
Rocamora’s and also Jeanselme’s experience, salvarsan has no
influence whatever on the Wassermann reaction in lepers.
Evidently neither Wassermann’s nor Eitner’s test can be
considered strictly specific for syphilis or leprosy. It remains,
however, to be seen whether a test could be devised in which by
using pure cultures of ‘Hansen’s bacillus’ a clear, specific, definite
reaction might be obtained.
Currie and Clegg approached the problem with the intention of
proving the identity of the acid-fast bacteria they had cultivated
from lepers, and after numerous experiments concluded that:
4 1. They were unable to differentiate by the method of Bordet and Gengou
the leprosy bacillus from certain other acid-fast mirco-organisms.
2. Extracts of certain acid-fast bacilli, other than B. leprae , will deflect the
complement when combined with leper’s serum.
3, They were able to produce specific agglutinins for B. leprae by injecting
a horse with the cultivated lepra bacilli.’
The explanation of these results is given by Much and his
co-workers, Hossli, Lescke and Deilmann, who have shown that all
acid-fast micro-organisms are more or less closely related and their
chemical constituents are similar. Much sums up his results in the
following words (l.c. 1912):
* We were able to show—chiefly by the reaction of immune bodies—a relation¬
ship between tubercle and lepra bacilli and other acid-fast bacilli, which is due to
inherent qualities common to all these organisms. Thus tuberculous and tubercle
immunised persons responded to the reaction of complement-fixation, not only
with tubercle, but also with other acid-fast bacilli, though not equally with all,
49
and with certain sorts more than with others. A table which I have prepared
shows the quantitative differences in these reactions, and according to this table
the lepra bacilli have the closest resemblance to the tubercle bacilli.
In the same way leprous sera react not only against lepra bacilli, which we
obtained from leprous tubercles, but they give also a specific reaction in the same
scale of relationship with tubercle and (other) non-pathogenic acid-fast bacilli.
These reactions are absolutely specific.’
Much’s observations and experiments show clearly that, as was
to be expected, the specific result of any serological test carried out
with acid-fast micro-organisms is apt to be masked by the inter¬
ference of a group-reaction. This is similar in its effects to that
observed on trying to differentiate from a serological standpoint
various bacteria of the typhoid or para-typhoid group. It is no
doubt also analogous to the results of precipitation tests when
being adapted to identify the blood of closely-related animals.
■'DOOOS^i
(•)(•)
• 9 ji
■)K K
'll '\i« '.»• ,*• '|i**
JuvAi
<ooooo®«®0oooo
••ooooo®#<§>® oooo
-00000®T»)®0GCC
‘OO.OO® ®*®®OGQO
% '(» ' t N 'i»i 144
Fig. i.
Diagram showing two complement fixation tests for the identification of acid-fast micro¬
organisms isolated from lepers.
Both serum and antigen are diluted in falling dosis and mixed in varying proportions.
Clear circles indicate complete haemolysis, dark circles absolute inhibition ; spots of varying
sizes correspond to the stages between haemolysis and inhibition.
Dark circles in the middle vertical line are controls of antigen, serum, complement.
H. R. Dean having shown that this difficulty can be obviated
by linking up antigen and antibody in varying dilutions, Bayon
carried out some experiments on the same lines with the intention of
getting rid of the group-reaction without having recourse to
Weichardt’s complicated technique. . These tests require the
greatest care, because the antibodies ot leprous sera are never very
strong, and dilutions capable of reacting above i /150 are the
exception. The antigen was prepared by alternately freezing and
rapidly thawing, and then shaking mechanically, various acid-fast
cultures in distilled water. The principal factor consisted in the
variation of the proportions of antigen and antibody, which can be
best explained by referring to the figure on p. 49, in which the dark
rings represent complete haemolysis and the clear rings complement-
absorption. It will be seen that in all the rows the leprous serum
was diluted in falling quantities, 1/5, 1/10, 1/20, etc. In the first
row the antigen was used undiluted, in the second 1/2, in the third
1 /4, and so forth. By this method not only a control was achieved
unobtainable by any other technique, but, moreover, very satisfac¬
tory and reliable results were arrived at. It was found possible to
distinguish, from a serological point of view, cultures of various
acid-fast bacteria, unless they were too similar or closely related,
e.g., bovine and human tubercle. In this fashion Kedrowsky’s
strain of leprosy was found to react specifically with the scrum of
lepers in a dilution of antigen which gave no result with Duval’s or
Rost’s micro-organisms.
This test can be applied to the diagnosis of leprosy, but
unfortunately, in common with many other laboratory methods, it
fails in those cases of early anaesthetic leprosy, which are the most
difficult to diagnose clinically.
In the instance illustrated in fig. 1 the serum of a nodular leper
was tried against antigens made with the leprosy cultures of
Kedrowsky and Duval. It will be seen that though Kedrowsky’s
micro-organism reacted even in a dilution of 1/4 with serum diluted
1/40, Duval’s bacterium ceased to give any absorption after mixing
1/2 with serum 1/5. These results have lately been confirmed by
Kritschewsky and Bierger, who however did not grade their
dilutions. In the second series of fig. 1 we see a test carried out
with the serum of a rabbit which had been repeatedly injected
intravenously with several slopes of Duval’s culture. The serum of
this animal was used against an antigen made with Duval’s 4 lepra,’
and another consisting of Moeller’s 'mist-bacillus’ isolated from
horse-dung. The result is that very little difference can be made
out, showing that Duval’s so-called 1 leprosy-culture ’ is very closely
related to a harmless, ubiquitous saprophyte.
These results were confirmed in course by the fact that the
etiological significance of this isolation had to be abandoned by its
author.
Moellers (1913) examined the sera from thirty-two lepers, using
as antigen the following products: Alttuberkulin Hochst, Perl-
suchttuberkulin Hochst, Tuberkelbazillenemulsion T.O.I., Neues
Tuberkulin T.R. He sums up his results in the following fashion:
(1) The serum of mixed and nodular lepers gave a positive
fixation of the complement in 95 to 100 per cent, of cases examined
when tested against various tuberculins. Anaesthetic varieties of
the disease reacted only in 25 per cent.
(2) In the serum of lepers the amboceptors for bacillary
emulsions are more developed than those for preparations made
from the cultural liquid.
(3) It is not possible to draw any conclusion as to the presence
of tuberculosis in a leper who shows the presence of 4 tuberculous
antibodies’ in his serum.
(4) The complement absorption properties of leprotic sera
tested with tubercular substances are in direct proportion to the
extent of mixed and nodular features in lepers. Accordingly it
appears that this inherent quality of the serum depends on the
greater or lesser spread of the leprotic lesions (Krankheitsherde);
this is confirmed by the observation that the reaction was negative
in arrested cases.
Kritschewsky and Bierger (1913) found that the serum of lepers
gave a definite positive complement fixation test with an antigen
made with Kedrowsky’s culture. Duval’s 1 lepra ’ did not react in
the same remarkable manner. A close similarity in relation to
serological tests was found to exist between the leprosy * bacillus ’
taken from tissues and that isolated by Kedrowsky. M. tuber¬
culosis appeared also to be closely related to the latter micro¬
organisms.
These authors conclude that Kedrowsky’s isolation is true
leprosy, and that Duval’s chromogenic culture is not etiologically
connected with the disease.
5 2
Gaucher and Abrami, Sugai and others, have tested the
agglutinating properties of lepers* sera towards Hansen’s ‘ bacillus *
separated from ground-up nodules. Such experiments are some¬
what vitiated from the fact that acid-fast rods are prone to show
spontaneous clumping.
Summing up these different independent observations it cannot
be definitely stated that it is not possible, with due care and
attention, to carry out serological tests showing the etiological
relationship of any given micro-organism to leprosy. In the case
of acid-fast bacteria the group-reaction must be obviated. The
difficulties are superior to those encountered with other germs; they
are, however, not insurmountable, as Harris and Langford are
inclined to believe.
Tuberculin and Similar Tests in Leprosy
Abraham, Goldschmidt, Babes and Kalindero, observed practi¬
cally at the same time, in 1891, that lepers react with a rise of
temperature to injections with ‘ Alt-tuberkulin.* Though this
observation was repeatedly confirmed, the tendency was to consider
that the reaction was due to a co-existent tuberculous infection.
Babes, however, examined seven lepers after death, who during life
had reacted to tuberculin, and found in four chronic tuberculosis
of the lungs, whilst the remaining three were without any signs of
infection with M. tuberculosis. Guinea-pig experiments were
negative.
Sir Malcolm Morris and Colcott Fox have observed very violent
reactions in anaesthetic lepers; in the former case bullae appeared
all over the arms.
Babes, having given considerable attention to the subject, is of
the opinion that some lepers react with quite a small dose, others
only after repeated larger injections. A general reaction takes
place after about twenty-four hours, in some instances, however,
after eight to ten, or even as short a time as only two hours, and as
a rule lasts longer than in the case of tuberculosis. The first can
be followed by a second or even third rise of temperature. Under
certain circumstances an extremely marked reaction may follow an
53
injection of less than one mmg., lasting several weeks and
endangering the life of the patient.
Jadassohn has, however, been unable to produce any reaction
with tuberculin in four cases he examined. It will be seen that the
evidence is contradictory, but if we take into consideration the
relative close relationship of leprosy and tuberculosis from many
points of view, it cannot appear improbable that lepers should react
to various tuberculin tests.
In previous communications Bayon was inclined to consider that
intra-dermal test after Mantoux’s method, carried out with a
cultural extract made from Kedrowsky’s strain of leprosy, might
have a specific and diagnostic value. Further investigations have,
however, shown that similar local and general reactions can be
brought about by the extracts of several acid-fast micro-organisms
such as M. phlei y Moellers ‘ mist-bacillus ’ and Duval’s so-called
‘ lepra.’ It is true that as a rule the reaction is not as marked with
the latter substances, but the difference is only one of degree.
On the other hand, lepers who do not react to a tuberculin test
may show very definite local reactions after intra-dermal injection
with one c.cm. of a io per cent, dilution of the extract from
Kedrowsky’s culture. This may consist of a copper-coloured patch
appearing at the site of injection or in more marked cases in the
production of blisters and infiltration (Plate V, figs. 3 and 4).
One patient at Robben Island, under observation of Dr. W. L.
Stuart, developed reddening and infiltration and a rise of tempera¬
ture to ioo° F. even after injection of minimal doses corresponding
to one c.cm. of a I /250 dilution of cultural extract. Other nodular
lepers again did not react even after injection of 3 c.cm. Both
extremes are exceptional; as a rule one or two c.cm. will be
followed by a rise of temperature and an intra-dermal injection
will result in varying degrees of discoloration. Pattison injected
ten lepers (four nerve cases, four mixed and two nodular) with
one c.cm. 1/10 dilution; all reacted either locally or generally. The
local reaction consisted usually of a dark patch at the site of
injection surrounded by a red zone, which might be swollen and
tender.
All these tests have, it is true, confirmatory value, but at the
present stage they are insufficient as a decisive help to diagnosis.
54
REFERENCES
Akxrbzrg, Almxvist und Jundell (1910). Weitere Beobachtungen uber Wassermanns Serum
Reaktion bei Lepra. Lepra, IX, 79.
Babes (*909). Sur des reactions reputes comme specifiqucs de la Wpre. Lepra III, 321.
- (1910). Uber spczifischc Reaktionen bei Lepra. Zcitsch. f. Imro. u. exp. Thcrapie,
VII, 578.
Babes ct Busila (1909). L’cxtrait des lcpromes gardes depuis des annccs dans Talcool comme
antigine lipreux. C.R. Soc. dc Biologic, LVII, 817.
- (1910). Etude sur le rapports qui existent entre les antigens et les anticorps
syphilitiques, tuberculeux et lipreux. C.R. Soc. dc Biologie, LVIII, 181.
Biehler and Eliasberg (1910). Komplementbindung bei Lepra mit leprosem Antigen.
Lepra, IX, 207.
Bruck und Gessner (1909). Uber Serumuntersuchungen bei Lepra. Berl. ldin. Wochen-
schrift, XLVI, 589.
Currie and Clegg (1912). Immunity. Lepra, XIII, 10-16.
Duval and Gurd (1911). Studies on the biology of and immunity against the bacillus of
leprosy. Arch, of Int. Medicine, VII, 230-245.
Ehlers et Bourret (1909). Reaction de Wasserman dans la lipre. Bull. Soc. de Path.
Exotique, 520.
Eitner (1906). Nachweis von Antikorpem im Serum eines Leprakranken mittels K.om pie-
mentablcnkung. Wiener klin. Wochenschrift, No. 5.
- (1908). Zur Frage der Anwendung der Komplementsbindungsreaktion auf Lepra.
Wiener klin. Wochenschrift, XXI, 729.
Eliasberg (1909). Komplementablenkung bei Lepra mit syphilitischem Antigen. Deutsche
med. Wochenschrift, XXXV, 1922.
Fox, H. (1910). The Wasserman and Noguchi Complement-fixation test in Leprosy. Am.
Journ. of Med. Science, May. Ref. Lepra, 1912, VIII, 107.
Frugoni (1909). Syphilis und Lepra. Arch. f. Dermat. u. Syph., XCV, 223.
Frugoni and Pisani (1909). Vielfache Bindungseigenschaften des Komplements einiger Sera
(Leprakranker) und ihre Bedeutung. Berl. klin. Wochenschrift, XLVI, 1531.
Gaucher et Abrami (1909). Le scrodiagnostique des formes atypiques de la lfpre. Lepra,
VIII, 152.
Jeanselme (1912). Cytologie ct serologic dc la l£prc. La Presse medicale, 27 Juillet.
Jundell, Almkwist und Sandmann (1908). Syphilisreaktion bei Lepra. Centralbl. f. inn.
Medizin, No. 48.
Kleinschmidt (1910). Bindung Komplementbinden der Antikorper durch Fette und Lipoid-
korper. Berl. klin. Wochenschrift, XLVII, 57.
Kritschewsky und Bierger (1913). Zur Frage uber das Verhaltnis des Bacillus leprae Hansen
zu einigen bie Lepra gezuchteten Mikro-organismen. Zeitsch. f. Hygiene, LXXIIT
509.
Lewin, A. (1911). Wasserman’s reaction in lepers. Russki Wratch. Ref. Lepra
XII, 229.
Meier, C. (1909). Serologische Untersuchungen bei Lepra. Lepra, III, 334.
MSllers, B. (1913). Serologische Untersuchungen bei Leprosen. Vcroffentl.
Stiftung, Heft VIII-IX, 122.
Much (1912). Serological and experimental Studies on Leprosy. Trans. Soc. Ti
V, « 75 -
- — (1912a). Uber Fettantikorper und ihre Bedeutung (mit besondercr Be
der Lepra). Beitrage zur Klinik d. Infekt. u. z. Immunitatsforschuncj
Kabitzsch.
ir
*
2 *-
r.'rf
Jr
Montesanto, Denis et Sotiriades (1910). La sero-reaction dc Wasserma
Upre. Presse med., 31 Aout.
55
Peyri-Rocamora (1912). Lc Salvamn dans la Wpre ; son influence sur le Wasserman dans
cette maladie. Lepra, XIII, 4-9.
Phonites et Michaxlides (1912). La sero-reaction de Wasscrmann et cuti-reaction de Pirquet
dans la Wpre. Lepra, XII, 207.
Recio, A. (1909). La reaction de Wassermann dans la Wprc. Sanidad y Beneficicncia Habana.
Sept. p. 292. Ref. Lepra, 1910, IX, 183.
Sf.rra (1939). La sicrodiagnosi di Wasserman nella Lepra. II Policlinico, No. 12, XVII.
Spindler (1912). Bcmerkungen uber.den Komplcmentgchalt und die Wasscrmannsche
Reaktion des Blutes Leproser. Dermatol. Zcntralblatt, XVI, No. 3.
Slattneano et Danielopolu (1908). Sur la presence d’anticorps specifiqucs dans le serum de
malades atteints de la lipre. Comptes Rendus Soc. de Biologie, 309.
- (1908a). Reaction de fixation dans la lipre en employant la tuberculine comme
antiginc. Ibidem, LXV.
- (1908b;. Reaction de fixation avec le serum et le liquide cephalo-rachidien des malades
atteints de lipre en presence de l’antigcne specifique syphilitique. C.R. Soc. de Biologie,
347-
SrcAi (1909). Zur klinisch-diagnostischcn Vcnvertung der Komplementsbimlungsmcthode
bei Lepra. Archiv. fiir Dcrmat. und Syph., XCV, 313.
Stepfenhagen (1910). Ubcr Komplementsbindungs reaktion bci Lepra. Bcrl. klin.
Wochenschrift, XLVII, 1362.
Thomsen und Bjarnhjedinson (1910). Untcrsuchungcn ubcr Komplcmcntbindung mit
dem Scrum Aussiitziger. Lepra, IV, 191.
Weciiselmann und Meier (1908). Wasscrmannsche Reaktion in einem Fallc von Lepra.
Deutsche med. Wochenschrift, XXXIV. 1341.
VIII. COMMUNICABILITY AND CONTAGIOUSNESS OF
LEPROSY
Exact and repeated observations carried out during the last
thirty years have definitely proved that leprosy is an infectious or
communicable disease. The 4 wisdom of centuries/ to quote
Sambon’s expression, had never doubted this fact. It is only due
to superficial clinical observers of the XIXth century that hereditary
and other irrelevant factors have lately been at all considered, even
for a short time, in connection with the disease.
The statement is often made that nothing is known about the
way in which leprosy is communicated from the diseased to the
healthy; but as a matter of fact, the question has been carefully
studied and the results are quite as definite, if not more so, than
any obtained in many other infectious complaints, such as measles
or scarlet fever.
Our knowledge of the contagiousness of leprosy is founded on
the following observations:
(i) That it is a disease due to a definite specific micro¬
organism which has only been found in lepers, their excreta or
immediate surroundings.
56
(2) That in Northern Germany, where leprosy had been
re-introduced from Russia in modem times, it was seen to have
spread very slowly and concentrically around the first imported
cases and their contacts.
(3) That on the island of Oesel, the population of which had
practically remained stationary during the last fifty-five years,
Lohk, Talwik, and Dehio were able to show that out of sixty-three
cases of leprosy only in eight could previous contact with lepers not
be traced.
(4) That with one exception (Da Costa) all cases, in which
contact cannot be proved, originate in countries or districts where
leprosy is fairly common; but that in the relatively rare instances
where leprosy has been contracted in countries in which the disease
is not indigenous, such as the United Kingdom, Southern Germany,
Holland, it is possible to prove the intimate contact with other
lepers, who in their turn had contracted the disease abroad.
(5) In countries where leprosy is rare, such as the Valais, the
Riviera, Alpes Maritimes, the disease is always found bound to
definite foci or families.
(6) That the countries which have carried out a thorough and
efficient system of segregation have been rewarded by a gradual
and constant diminution of the disease. (Norway, Iceland,
Germany, Sandwich Islands, Philippine Islands.)
(7) That where segregation has been abandoned, or loosely
carried out, or not enforced at all, the scourge has usually attacked
an ever increasing number of individuals. (India, Basutoland,
Dutch Indies).
The grounds on which the contagiousness is denied are, I
believe, the following:
(1) That medical men and attendants never contract the disease
in leper-asylums.
This statement is not quite correct. Infection under these
circumstances is not frequent, but still several cases can be brought
forward showing that attendants and medical men do contract the
disease. In South Africa the Medical Superintendent of a Leper
Asylum contracted leprosy. Further instances of professional
infection have been published by Ehlers, Vidal, Jeanselme,
Nicolas, and others.
57
In one case of which I know, a missionary to lepers, trusting no
doubt to the low degree of infectivity of the disease, allowed his
little girl to play with leper children. She developed the malady
whilst at school in Europe.
Published and unpublished cases of infection among religious
persons who have attended lepers are also known to exist. (Three
priests at Molokai, Father Boglioli in New Orleans, two nuns, a
missionary, a lady missionary.)
On similar grounds it has been attempted to deny the
infectiousness of tuberculosis, the deduction being based on
statistics showing the low ratio of marital and professional
infection.
(2) Because the disease cannot be transmitted to animals, and
inoculation experiments on human beings have failed.
This also is not quite correct. The first part of this argument
would have allowed us fifteen years ago to deny the contagiousness
of syphilis. As a matter of* fact, leprosy is transmissible to
laboratory animals such as the rabbit or rat, as repeated experi¬
ments have shown. That this possibility is not more generally
recognised is due to the fact that the lesions may be localised and
fail to develop at all in a very great proportion of experiments, and
often resembles tuberculosis to a certain extent.
As to the inoculations which Danielssen, Profeta, Cagnina,
Bargilli, carried out on themselves and others, the results, it is
true, were negative, but in view of the knowledge we have lately
acquired of the bionomics of the leprosy micro-organism, they have
only proved that experimental inoculation with small quantities of
leprous material is incapable of producing the disease in the human
being in every case.
In Aiming's well-known case of the convict Keanu who was
pardoned on condition he allowed himself to be inoculated with
leprosy, the disease did develop, but the experiment is somewhat
spoiled by the fact that the man had lepers among his relatives and
was in contact with them before and after inoculation.
(3) That in Norway leprosy diminished considerably in the
course of sixty years, but that only about a quarter of the lepers
were ever segregated at one time.
This is not quite correct; it is true that only a small proportion
58
of all the lepers known to exist were placed under strict segregation,
that is, in special asylums, but these were the cases unable, incapable
or unwilling to carry out segregation at home. All the rest were
avoiding at home all contact with their relations, etc., under
medical supervision. In this sense the system was one of efficient
universal segregation.
A further similar argument, often brought forward by non-
contagionists, is that leprosy disappeared in the Middle Ages from
England, France, Germany, etc., without strict measures of
segregation having been resorted to.
It appears to me that the measures adopted to prevent the spread
of the disease were extremely severe. Lepers were not allowed into
churches or market places, had to carry a distinctive dress, and had
to make their presence known by a bell or clapper. In a few words
the intimate contact between the healthy and the diseased, which
gives rise to the most favourable conditions for transmission, was
most radically avoided. I also believe that plague directly and
indirectly swept away a great proportion of the leper population,
as they were feeble, and during times of epidemics and commotion
would receive no alms. This observation has been confirmed to a
certain extent in India in modem times.
It will be seen that non-contagionist arguments are founded
mostly on negative evidence. Positive figures bearing on the
contagiousness of leprosy are available from Norway, Japan, and
the Sandwich Islands, and they unanimously point to the same
moral.
Kitasato’s statistics from Japan show that children of lepers
become leprous in a proportion of only 7*95 per cent, of the total.
Matrimonial infection was proven in 3 8 per cent, cases, whilst
persons living under the same roof contracted leprosy in a propor¬
tion of only 27 per cent. Brothers and sisters infect each other in
a ratio of 4*2 per cent. These figures may need correcting
according to the latest statistical methods. However, they roughly
correspond to the experience gained in Norway where Sand and
Lies’ figures differ somewhat, but show that the children of a
leprous mother are more frequently infected than those whose father
alone is diseased. This proves that the more intimate contact
between mother and child leads to a greater percentage of
acquisition of the scourge.
59
Sand’s statistics from Norway show that in 357 married couples
in whicfrthe father alone was a leper, 1,241 children were born, of
which 63, equal to a proportion of 4*9 per cent., became lepers.
In 138 other married couples observed, the mother only was a leper;
of 533 children born of these unions 56, or 10*5 per cent., developed
leprosy. In 17 couples both parents were diseased; of the resulting
63 children 8, or 127 per cent., became infected.
Lies’ figures from South-Western Norway give somewhat
similar results. 230 married couples in which the father only was
a leper had 769 children, of which 79, or 10*2 per cent., contracted
the disease. In 223 married couples the mother only was leprous;
of 648 children, 106 of these, otherwise 16 36 per cent., became
lepers. In 28 instances both parents were lepers; out of 79 children
29 fell victims to leprosy, that is, 39*19 per cent.
McCoy and Goodhues’ observations have been made in the
Sandwich Islands, and relate mostly to cases of infection noted
among the kokuas or voluntary helpers of the lepers at the Molokai
settlement. Their conclusions show that :
(1) Of 119 men, practically all Hawaiians or persons of mixed
Hawaiian blood, living in the same house with the lepers, five, i.e.,
4*2 per cent., developed leprosy.
(2) Of 106 women, practically all Hawaiians or persons of
mixed Hawaiian blood, living in the same house with lepers, five,
i.e., 4 7 per cent., developed leprosy.
(3) Of 12 women, all Caucasians, who lived in such contact
with lepers as is necessary in administering to their bodily and
spiritual wants, none developed the disease.
(4) Of 23 men living under the same conditions in contact with
lepers, three, i.e., 13 per cent., developed the disease.
(5) The shortest period in which the disease developed after
the person entered the settlement was three years (two cases), and
the longest seventeen years.
In a report made in 1886 it is asserted that of 178 kokuas, 17
became lepers in a year. In a later report, made in 1888, it is
remarked that of 66 kokuas examined 23 were found to have become
lepers.
Such a condition of affairs does not exist at the present time at
Molokai, and the change for the better is no doubt due to the great
6o
improvements made in the sanitary conditions of the settlement in
recent years. *
Of course it should not be forgotten that, as the kokuas are
drawn from a population in which leprosy is not very uncommon, it
is quite possible that a certain number of cases came to the settle¬
ment in a stage of incubation.
Hollman examined carefully the conditions affecting the develop¬
ment of leprosy in the children of lepers at the Molokai Settlement,
and came to the following conclusions:
(1) It is shown that 40 per cent, of the children born of parents
of whom one or both were lepers died under one year.
(2) 32 per cent, of the males who were exposed ten or more
years developed leprosy.
(3) 4 per cent, of the females whose average time of exposure
was less than five years developed leprosy.
(4) 10 per cent, of the males exposed for more than seven years
developed the disease.
( 5 ) *3 P er cent, of the females exposed from one to seventeen
years, and under observation seven or more years, became lepers.
(6) The average time of exposure of the cases which developed
leprosy was five years.
Accordingly, the danger of contracting leprosy for children
born of leprous parents increases with the length of exposure.
Incidentally these investigations show that heredity does not
play any important role in the causation of leprosy. Also, if it
did, the disease would soon die out in any country.
The figures and conclusions show so clearly that contagion or
infection through immediate contact is the usual mode of
communication, that it appears rather far-fetched to seek an insect
carrier of the scourge.
As a matter of fact, all experiments to prove this mode of
transmission have so far failed, though it appears quite probable
that the common house-fly can suck up the germs of the disease from
open sores, carry them about for several days, and disseminate
them in such a fashion.
The horrid sight of flies swarming and hovering over the
purulent sores and round the nostrils of leprous beggars is well
known to the traveller in eastern countries.
6i
Graham-Smith, having shown that house-flies can harbour the
bacteria of tuberculosis for twelve days or more, it seemed probable
that the micro-organism of leprosy would show an equally long
permanence in the fly-intestine.
Leboeuf examined numerous specimens of Musca domestica
caught on the sores of lepers, in the wards, and in houses not
further than 150 metres from the hospital.
He found leprotic ‘ globi' in the intestines of flies captured and
kept for twenty-four hours, and acid-fast rods in flies thirty-six
hours after feeding. His conclusions are that :
(1) Musca domestica can absorb enormous numbers of Hansen’s
‘bacilli' by nourishing itself on sores containing these germs. .
(2) The ‘bacilli' can be found in abundance and, apparently,
excellent condition, in the excreta of the infected house-flies.
(3) It does not seem that multiplication takes place in the
digestive tract of Musca domestica , but in any case there are no
signs of degeneration.
(4) Musca domestica possibly plays an important part in the
dissemination of leprosy by depositing its excrements on the mucous
membranes or small abrasions of the skin of healthy people living
in the immediate vicinity of lepers whose sores contain ‘ bacilli.'
It will be seen that in any case the fly does not do more than
eventually disseminate the micro-organisms it has ingested, in a
similar fashion to flies disseminating typhoid.
As far as our knowledge goes, no insect plays a real role of
transmission in any bacterial disease. Moreover, transmission
implies a more or less complicated developmental cycle in the body
of the intermediate host, after which a protozoon can be inoculated
by the proboscis of a biting or stinging insect. With bacteria a
contaminatory communication through the faeces or by the regurgi¬
tation of the crop contents takes place. This is the case in bubonic
plague or typhoid.
The fly is eminently adapted for a contaminatory or mechanical
method of dissemination, but the difficulties inherent to the
communication of leprosy to animals will render experimental work
in this direction very difficult to accomplish.
It will be seen that the intimate personal contact, as found
between a child and its mother, gives the most favourable circum¬
stances for the acquisition of leprosy.
hi
In South Africa, where the use of coloured and native servants
and nurses for children is very widespread, it is not astonishing to
find that about 40 to 50 per cent, of the white (Caucasian) patients
under segregation cannot tell where they have contracted leprosy.
Leprosy is quite widespread in the native population. In single
instances, after a great deal of investigation, I was able to find out
that the disease could be traced to a coloured nurse or servant who
had evident signs of leprosy or eventually had been segregated in
some asylum of the Union, the disease developing in the child many
years afterwards.
Native labourers suffering from leprosy appear to be the source
of contagion in many other cases. The Cape of Good Hope
Leprosy Commission of 1895, having given careful attention to this
matter, quotes several observations which support these views:
‘ An individual may be leprous for many months, and it may be for a year
or longer without showing outward signs of leprosy, and during that time may
have communicated the disease to several without being aware of the fact that
he is a leper himself. Hence arises the difficulty of knowing how and when the
healthy may have been in contact with the contaminated. It is self-evident
that the length of the period of incubation, and the possibility that during the
period of incubation one individual may communicate the disease to another,
greatly add to the difficulty of proving the source of contagion. Hence one may
readily understand that statement of many lepers, that they had never been in
close contact with a leper, or had never seen a leper, and consequently do not
know how they got the disease. In many cases, however, possibly in by far the
greater majority, it may be shown that at some period or other those who have
acquired the disease have been in contact with a person or persons suffering from
the disease ; and even where direct contact cannot be proved, indirect means
of acquiring leprosy, such as the use of the same bedding, or articles of clothing,
or utensils, or a pipe, or implements used by a leper, or living in a room occupied
by a leper, or attending to a leper, may be proved to have been present at some
time or other in the life history of those who have contracted the disease.
In the evidence given before your Commission, Dr. A. J. J. Simons, of
Malmesbury, who, in the course of an extensive practice reaching over a period
of years, had many opportunities of observing the disease, gives a very instructive
history of the course of leprosy on a certain farm in that division. In the case
under consideration the leprosy was first noticed by him in a coloured man. a
bastard Hottentot; next a little girl, the farmers daughter got the disease ;
next another coloured man on the farm, a wagon driver who used to be great
friends with the bastard Hottentot became diseased; after the wagon driver
his master, the farmer, became leprous; next a girl who came to the farm to attend
the farmer’s daughter and nurse her ; and finally the farmer’s niece who frequently
visited the farm, spent the day there and was generally intimate with the leper
daughter of the farmer. Everybody who is acquainted with the patriarchal
63
mode of life of the South African farmers, and who knows the habits of our rural
population, will admit that the account given by Dr. Simons is a history typical
of what happens in every country district in South Africa generally.’
* Dr. Nieuwoudt, Darling, records the following cases: (i) A patient, in
whose family, apparently, the disease was not present, was waited on by a leprous
servant, and developed the disease in a mixed form. Later on a boy who attended
on him, etc., got the disease and died ; later on the gentleman’s daughter developed
the disease in the tubercular form and is still here, though her father has since
died. (2) An old man had the disease in whose family the disease did not exist,
so far as known. His brother’s son, who wore the same hat of his, got the disease ;
and another man outside this family, and of a clean family, in whose bed the
patient slept, also developed the disease.’
‘ Dr. Newnham, Aliwal, knows a case where a farmer was clearly infected
by his native coachman who had been long a leper. Master and servant when
travelling together used the same horn drinking cup.’
‘ Dr. Vanes, M.L.A., of Humansdorp, writes: “ In a family living at Patentie
there were four children affected, the parents being healthy.” With regard to
these last cases it must be noted that many examples may be cited, which prove
that children get the disease before the parents. Such cases appear to your
Commission to plead powerfully in favour of the contagiousness of leprosy and
as powerfully against its heredity. More especially in those cases in which no
remote family taint can be shown.
Dr. Barry, of Bedford, quotes the case of a Dutch farmer, V.H., who states
that he got the disease through handling a spade used by a leper Hottentot.’
These observations might possibly be set aside by the hyper¬
critical if they did not agree in a striking fashion with the inferences
deduced from the investigation of the spread of leprosy in many
other parts of the world.
We know that at the present moment about twenty-five to fifty
lepers or more are living in England : and yet of these only one has
acquired the disease in the United Kingdom (mother and father
were lepers). The simple precautions these unfortunates are able
to take to keep themselves separated from their families have been
sufficient to prevent contagion.
As a contrast we have India, where the last census appears to
show an increase of lepers in the last ten years from 100,000 to
110,000.
The modern medical eye looks, therefore, upon leprosy as a
disease which is definitely contagious, but to a very slight degree
6 4
under proper sanitary conditions. In situations where hygienic
precautions are defective and the contact between the diseased and
the healthy is unnecessarily immediate, where a leper is obliged to
sleep in the same bed with other members of the family, and
personal cleanliness apt to be in abeyance, the danger of contagion
is certainly present to a markedly increased extent.
REFERENCES
Dehio, K. (1910). Uber die Verbreitungsweise der Lepra. Lepra XI, 16-24.
Hollmann, H. T. (1910). Heredity versus Environment in leprosy. Public Health Bulletin
No. 39, XI, 43-49.
Honeij and Parker (1914)* Leprosy : flies in relation to the transmission of the disease (a
preliminary note). Joum. of Med. Research, XXX, 127.
Kjtasato, — (1910)- Die Lepra in Japan. Lepra, X, 144-151.
Leboeup, A. (1912). Dissemination du bacille de Hansen par la mouche domestique. Bull.
Soc. Path. Exotique, p. 860. Ref. Lepra, XIV, pp. 119-124.
Lie, H. P. (1911). Statistisches uber Lepra. Archiv fur Derm, und Syphilis, CX, 473-486.
McCoy and Goodhue (1913). The danger of association with lepers at the Molokai Settle¬
ment. Public Health Bulletin No. 61, XX, 7-10.
Sand, A. (1903). Beobachtungen uber Lepra. Lepra, III, 7-16.
- (1910). Gcschieht die Ansteckung der Lepra duch unmittelbarc Ubertragung ?
Lepra, XI, 39-46.
IX. EARLY AND DIFFERENTIAL DIAGNOSIS. PROGNOSIS
The early recognition of leprosy presents difficulties commen¬
surate with its importance; for at times the symptoms may be so
vague and indefinite that in the absence of conclusive microscopical
evidence a correct diagnosis can only be made by a process of
exclusion. Cases in which nodules containing numerous acid-fast
rods, or in which typical maculae accompanied by anaesthesia or
even contractures can be detected, cannot generally be considered
as early conditions. Moreover, these ought to be diagnosable by
someone who has a practical knowledge of the disease. It is in
connection with slight and evanescent symptoms that serious
difficulties arise, for such indefinite appearances are generally the
only ones noticeable during the prodromal stage.
Lepers with very slight and atypical lesions are not necessarily
early cases from the chronological point of view, because the slight
areas of anaesthesia or discoloration may have existed unnoticed
for years.
Sir Patrick Manson mentions the case of a gentleman who
developed leprosy live years after noting in Ins diary a long series
of headaches, transient fever attacks and progressive deterioration
of health and vigour. These prodromal symptoms were followed
three years afterwards by profuse perspiration which occurred
without obvious cause. Such carefully observed cases are not
frequent, and in view ot the abnormal length of the period of
incubation of leprosy it is often impossible for the patient to
remember the occurrence of slight indefinite prodromal symptoms.
What in many cases is apparently the onset of the disease is
but the more marked appearance of symptoms which have been
present for a long time. Correspondingly very slight evidences of
leprosy may be the traces of much more apparent previous patho¬
logical processes. It is accordingly often practically impossible to
say whether we are dealing with an early case, though the symptoms
may not be conclusive.
It should not be taken for granted that any case showing
leprotic symptoms necessarily will progress and become irrecog-
nisable through deformities and mutilations because leprosy may
get arrested in the incipient stage.
A disease with such multiform variety of clinical features is
naturally often extremely difficult to recognise in its first stages,
not only because of its tendency to produce symptoms which are
not typical for any single condition and common to several ailments,
but also because of the corresponding variety of initial appearances.
As a rule anaesthetic cases begin with a peculiar numbness in a
digit, preferably the little finger of one or both hands. This
numbness the patients compare to feeling that their fingers are
covered with leather. On plunging the hand in warm water a
peculiar sensation is noticed in so far as the finger or fingers affected
feel the pain caused by the heat to a much lesser degree. The same
symptoms may first show themselves in a little toe. On observation
it may be seen that the fingers affected are glossier, more shiny than
the corresponding or other fingers of the same hand. On taking
exercise in warm weather it will be noticed that these areas are much
less apt to perspire. Contemporary with these anaesthetic symptoms
maculae or discoloration may be present or not in other parts of
the body. Under some circumstances the maculae can be so slight
66
as to amount to little more than a peculiar discoloration very similar
to cutis marmorata or mottled skin due to effects of cold, or they
may take the shape of slight red lines which may form a closed
figure. In course of time a case like this becomes a definite maculo-
anaesthetic leper, or develops nodular features. A third alternative
is that the symptoms may slowly but surely disappear, leaving
hardly any trace.
The insensibility to heat and cold may be accompanied or
heralded by formication and shooting neuralgic pains in the limbs
affected. Profuse sweating precedes often the anhydrosis.
Pigmented spots may appear on various parts of the body,
varying in colour from a light lemon yellow to a dark dirty brown.
On looking at the spots under various angles of light incidence it
will be noticed that the skin does not reflect the light everywhere in
the same fashion, but is rougher in parts and appears covered with
a slight farinaceous deposit which consists of desquamating
superficial epithelial cells; the surface is intersected with very fine
wrinkles, and has a dry atrophied appearance.
Another early stage consists in puffiness and infiltration round
the alae nasi, or thickening of the lobes of the ears and loss of the
eyebrows, accompanied often by intensive irritation and itching of
the neck and back of the scalp of the head. In such instances
epistaxis and dryness of the nostrils may be followed by a
persistent cold in the nose.
A further early stage may be shown by a small pimple or blind
boil, appearing in some part of the body, generally the face, this
being followed by an outcrop of typical small nodules, superficial
or subcutaneous. Under certain circumstances some sore or wound
may be found to heal with extreme difficulty, or a slight abrasion
may turn into a chronic ulcer; these have often been considered to
form the initial sore or lesion of leprosy. They are at times,
however, but a symptom of an infection which is already fully
developed.
In dealing with such extremely early cases one should never fail
to map out the extent and measure the intensity of the paraesthetic
symptoms, remembering that in the beginning maculae are usually
hyperaesthetic; pin-pricks, etc., are painful which under ordinary
circumstances would cause hardly any discomfort; the sense of
6?
touch can be unimpaired. Special value is to be laid on the faculty
of recognising blunt and sharp (a penholder or a sharpened lead
pencil will do for the purpose) and in each case a trial with two test
tubes filled with warm (50°-6o° C.) water, and cold water should be
used to discover the patient’s sensibility in distinguishing hot and
cold in the affected areas. Slight muscular atrophy of the interossei
should be looked for.
In all doubtful cases a microscopical examination of maculae or
discolorations should be undertaken, though the interpretation of
any histological lesions observed is a matter requiring expert
knowledge. Needless to say, acid-fast rods should be searched for
in the nasal mucous membrane; they may be present for a short
time and then disappear for longer or shorter intervals.
Small nodules can be grouped up and treated with Anti-formin,
centrifuged and the sediment examined for bundles of acid-fast
rods. This method should, however, be only used in conjunction
with an histological examination, because single acid-fast micro¬
organisms, without the lesions described elsewhere, are not sufficient
to diagnose leprosy.
The fusiform thickening of the ulnar or other nerves, which is
one of the cardinal symptoms of anaesthetic leprosy, as a rule does
not come into practical consideration as a help in the early
diagnosis of leprosy, because in the initial stages it is not usually
present to a sufficient degree to render its detection easy. Moreover,
such swellings are often deeply seated, and therefore not accessible
in clinical examination.
Differential Diagnosis
Apart from the histological and serological features which have
already been discussed, the differentiation of leprosy from other
skin lesions, which are somewhat similar in appearance, is generally
based on the assumption that leprosy is an extremely slow and
intractable disease, and that its manifestations are essentially
indolent in character.
Much has been written on the difficulty of distinguishing leprosy
from syphilis. There can be no gainsaying that in very rare
instances—a case has been illustrated by Graham Little—a
68
momentary confusion can arise, but a practical knowledge of both
complaints soon shows that they must be quite different in their
naked eye appearances.
Syphilis and yaws as a rule produce, in all stages, lesions which
are more vascular, angrier, with a more active and proliferating
appearance than leprosy, which is an eminently indolent, torpid,
slow pathological process.
This difference is also applicable to the scars left by ulcers in
these diseases. Syphilitic scars are usually rough, with numerous
small fleshy warts and excrescences, and their margins are irregular
with numerous small pockets and indentations. At times peculiar
minute bridges of tissue can be observed under which a sound or
needle can be passed. All these appearances are the result of a
process of active proliferation and production of cicatricial tissue.
Leprosy leaves scars which are smooth, shiny, and usually flush
with or gradually merging into the surrounding healthy skin.
Accordingly any circinate macular efflorescences in syphilis and
yaws should be more raised, redder, bleed more easily than a
corresponding leprotic manifestation. In addition they should
respond to mercury or salvarsan.
Practically every macular leper shows at least incapacity of
distinguishing blunt from sharp and hot from cold (therman-
aesthesia) in some area of his body. Without this crucial test it is
not advisable to diagnose leprosy, however typical the lesions may
appear to the naked eye, unless the opinion can be confirmed by the
microscope.
Leucoderma in the negro and dark-skinned races is often
mistaken for leprosy. Here again the lack of anaesthetic or
thermanaesthetic symptoms, the uniformity in hue and spread of
the white spots, ought to facilitate a decision.
Lupus and tuberculosis of the skin are the pathological processes
whose manifestations may cause considerable difficulty. However,
lupus is rarely multiple, has an early tendency to break down and
ulcerate, forming scars at its periphery. Lupus is more friable and
bleeds less easily, and though more destructive than leprosy is
usually somewhat more amenable to local treatment.
Pellagra ought not to be confused with leprosy, and vice-versa.
Pellagra is symmetrical, strictly confined to parts generally exposed
69
to the action of the sun: there are no marked disturbances of
sensibility in the areas affected, and the lesions are much more
uniform and regular than is usually the case with lepra. The edges
are not indented or irregular. In doubtful cases the microscope
should decide.
The ringed circinate maculae seen in Caucasians in early stages
of trypanosomiasis are not easily distinguished from some phases
of early macular leprosy; in fact in one case to my knowledge the
presence of the diagnosis was only cleared up by finding trypano¬
somes in the blood. Leprotic lesions are, however, more permanent
and are not as fleeting as the similar eruption in trypanosome
disease (‘ trypanides ’).
Syringomyelia, especially in its initial stages, is extremely
difficult to distinguish from lepra, unless oculo-pupillar paralysis
(consisting in myosis, diminution of the eyelid aperture and
decreased prominence of the eyeball, Moritz) is present in the
former eventuality, or typical maculae can be detected in the latter.
A search for clumped acid-fast rods should be made. Early leprosy,
moreover, does not often show such generalised lesions as
syringomyelia.
In advanced cases, symptoms on the part of the medulla
oblongata, scoliosis, and the absence or presence of thickened
nerves, ought to help the diagnosis.
The absence or presence of thermana esthetic and analgetic
symptoms should be carefully investigated when dealing with
coloured races, for numerous skin diseases due to fungi cause
lesions which resemble leprosy in quite a disconcerting fashion.
Here, again, microscopical evidence should not be missed.
In conclusion, it need only be added that in many instances the
locality from which a patient comes, etc., the possibility or
probability of contagion, are facts which, though they cannot carry
conviction in every case, still may on the other hand simplify the
diagnosis. It will be seen that very often recourse will have to be
made to trained microscopical and bacteriological examination of
excised fragments of skin.
The extreme variability of all the initial features of leprosy
and the chronic course of the complaint, allow only a most guarded
prognosis to be stated in any single case of the disease. It cannot
70
be denied that, given skilful medical attendance and healthy
surroundings, even this most dreaded complaint can be held in
check for comparatively long periods. It is true that little can be
done for advanced cases, except from the standpoint of surgery.
Early cases, however, will show but little impairment of their
general health under favourable conditions and arc quite capable
of living a useful life for years and years.
From what has been said in connection with early diagnosis, it
is evident that it is not safe to predict in the early stages what
course leprosy is going to take; as a rule, cases which have begun
with indefinite anaesthetic symptoms, last longer than those whose
nodules have appeared in batches or have shown an extensive
appearance of simultaneous patches.
The extensive mutilations of many anaesthetic lepers are but
too often due to lack of attention to trifling casual wounds and
burns.
X. THE TREATMENT OF LEPROSY
Not only nearly every imaginable drug, but in addition poisons
and venoms, have been tried on lepers with the intention of
discovering a cure, but this notwithstanding we have yet to find a
prompt and sure method of treatment.
The present position can, however, be summed up in the words
that even the most useful therapeutic substance still requires the help
of an early diagnosis and proper hygienic conditions; but that at
no time has the outlook regarding the treatment of leprosy been so
hopeful as at the present moment.
In its initial stages leprosy affects the general well-being and
appearance of an individual to such a slight extent that any thera¬
peutic effort which succeeds in arresting the disease in its early
stages in a fair proportion of cases would practically amount to a
cure.
Spontaneous remissions of the disease, and spontaneous
apparent ‘cures* take place, however, in a small proportion of
lepers, and can last for several years, in some instances as long as
15 years or more, after which the disease may again become virulent
and rapidly carry its victim to the grave.
7 1
Therefore any drug or method of treatment must stand the test
of time—five years or more—and have been carried out on a
sufficient number of patients.
Without taking into consideration these important factors it is
absolutely irresponsible to speak of cure in a chronic, slow,
intractable malady such as leprosy.
A review of all therapeutic attempts would fill a volume by
itself. I may, however, be allowed to give the results of the
experiments made at Robben Island, and in South Africa generally.
For all advanced nodular stages Chaulmugra oil, or better still
its refined constituent 4 anti-leprol,’ injected intra-muscularly
3-5 c.cm. at a time, is still the best palliative I know of. The
injections should be repeated every three days and the course last
five months or more if the patient can stand it, because at times the
injections may become very painful. Chaulmugra oil and
Antileprol can also be given internally in small capsules.
Antileprol is decidedly preferable for the latter mode of
treatment, as it does not cause the gastric disturbances which are
the outstanding feature of the unrefined oil. Doses varying between
15 m. and ten times that quantity can be taken daily.
Chaulmugra oil is a product of the seeds of the Taraktogenos
kurzii , whilst the oil extracted from Gynocardia odorata ought to
be called false chaulmugra (D. Hooper; Article on Taraktogenos
kurzii in ‘The Agricultural Ledger/ 1905, No. 5). It appears that
substitution and adulteration of this oil is extensively practised,
owing to the difficulty of keeping up a regular supply of the seeds
of the Taraktogenos kurzii . Further particulars relative to this
important subject are to be found in the paper by Hooper, which
no doubt explains the variable results obtained in treating lepers
with chaulmugra oils from different sources.
Brocq and Pomaret mix 70 parts of chaulmugra oil with 30 of
eucalyptus oil, and find that the injections of this product are much
less painful than the pure oil.
Early macular cases appear to respond better to other forms of
treatment. A cultural extract has been prepared from Kedrowsky’s
isolation of the micro-organism of leprosy on similar lines to Koch’s
Alt-Tuberkulin. It contains accordingly the water-soluble toxic
constituents of the bacillus, which, however, in the case of Hansen’s
7 *
germ are very feeble indeed, so that much higher concentrations
have to be injected than is the case in the analogous treatment of
tuberculosis by tuberculine.
Dr. MacLeod of Charing Cross Hospital, and Dr. T. S. Davies,
Resident Medical Officer of the Pretoria Asylum, have made use of
this substance on single chosen cases. The results in both instances
have been very satisfactory. The maculae or spots have slowly
faded away.
Dr. MacLeod's patient, a young lady from the West Indies,
has improved to such an extent that the disease is no more
diagnosable. I am authorised to say that other therapeutic efforts
failed to bring about any change for the better, and that the
patient has now been observed for over two years without any
remission taking place. On the contrary a further subjective
improvement has so far taken place that muscular pains which were
present in the arm affected have disappeared.
Dr. Sidney Davies's first patient, a little girl of years, was
covered with dusky red patches all over her body. These faded
away in the course of treatment extended over eighteen months,
leaving only a slight discoloration (PI. VI, fig. 6).
Later, six other lepers were injected with the same cultural
extract at Pretoria, with the result that a remarkable improvement
was brought about in two (this includes the patient already
mentioned), marked improvement became noticeable in three, and
slight improvement in the remaining two.
The injections are being continued, and at the time of writing
further progress has been made, that is to say, that the maculae
have continued fading and in some cases have disappeared, leaving
only a trace of faint discoloration. Details will be published in
due course.
It now remains to be seen whether these results will stand the
test of time, and whether the same amelioration takes place in
larger experimental series.
This method of treatment does not appear to be of real
advantage in advanced nodular cases. The injection of one or two
cubic centimetres of the cultural extract causes a rise in temperature
which is accompanied by the appearance of bacilli in the blood.
This may possibly lead to dissemination of the germs of the
73
disease. If the individual affected is young and otherwise healthy,
and the leprotic symptoms not marked, a course of injections
beginning with one cubic centimetre hypodermically every three days
may be distinctly beneficial. The treatment should be continued
for some time even after the maculae have disappeared.
The surgical treatment of leprosy requires most careful and
constant attention. Fingers, toes, hands and feet are constantly
getting septic, and prompt surgical intervention can really work
marvels. Eye complaints, which are exceedingly frequent among
lepers, are amenable to a certain extent to operative procedures, but
relapses often follow even after the most skilful intervention.
REFERENCES
Brocq ct Pomaret (i913). Nouveau produit injectable pour le traitemcnt de la lepre. Ref.
Lepra, XIV, 124.
Davies, T. S. (1914). Further notes on the specific treatment of leprosy by a cultural extract.
S. Afr. Med. Record, XII, 77-8.
MacLeod, J. M. H. (1912). Case of maculo-anaesthedc leprosy in a woman aged 25. Proc.
Roy. Soc. Medicine, V, 142-4.
Serra, A. (1913). L’antileprol dans le traitement de la lepre. Lepra, XIV, 63-9.
XI. PREVENTION, ASYLUMS AND HOME-SEGREGATION.
SETTLEMENTS AND LEPER COLONIES
The trend of all recent research has been to prove conclusively
that though leprosy is communicable from the diseased to the
healthy by direct and indirect contact, still relatively simple
precautions are sufficient to reduce the peril of infection to a
considerable extent. Thus the probability of the scourge rapidly
spreading and giving rise to an extensive epidemic is certainly
remote. On the other hand, if hygienic precautions are neglected,
the danger for the immediate surrounding and family of a leper is
by no means to be despised, in view of the gravity of the disease.
The fortitude with which some nations view the slow but
continuous increase of leprosy in their midst deserves to . be
explained, at least to obviate its being misunderstood for some less
admirable virtue.
If the news spread that robbers or thugs or wild tribesmen were
killing, torturing or maiming about five hundred or a thousand
74
people a year, I have no doubt that expeditionary corps, the police,
and all the full weight of official indignation, would be employed
to crush the criminals out of existence, and quite rightly so. On
the other hand, an acid-fast micro-organism, against which we have
hardly any chance of defending ourselves unless we are armed with
the sharpest weapons of preventive medicine, is allowed to corrupt
the bodies and souls of hundreds and thousands of new victims
yearly without a single protest being heard. This is due to the
fact that a biological mode of thought has not yet permeated the
body politic, and that accordingly it is not yet sufficiently under¬
stood that public health means public wealth and individual health.
We know of no other method of eradicating leprosy from a country
than by segregation. All the critics of this method of dealing with
the disease have no other practicable suggestion to make. It is
true that by improving the hygienic conditions of the homes and
lives of the poorer population, by carrying through an efficient and
widespread sanitary inspection, leprosy could be slowly and
definitely stamped out; but to deal with the scourge promptly
nothing else can be suggested but isolation. The only argument that
can be brought to bear against segregation is on the score of expense.
In some instances parsimony and reluctance to invest money in
public health matters have been veiled by the sentiment that it
would be cruel to separate a leper from his family and his friends.
Two facts are, however, overlooked. One is that when an
individual develops leprosy in a severe and noticeable form, he
need not trouble about his friends: they leave him of their own
accord. The other is that consideration of the feelings of one may
mean the misery of the whole family. In South Africa one leper
infected in the course of time, directly and indirectly, sixteen other
members of his family.
It is erroneous to believe that all lepers being segregated are
compulsorily detained, however penurious may be the conditions
offered by an asylum. Many are but too glad to find a refuge, a
shelter from the dreadful conditions and inhuman circumstances
under which they had to exist in the outer world.
The new introduction of a scheme of segregation requires careful
preparation, because experience teaches us that the mere building
of asylums is not sufficient to eradicate successfully the disease.
75
Efficient isolation presupposes adequate, skilful, sympathetic
medical attendance in asylums, and when such is forthcoming many
of the terrors of segregation disappear. Even in leprosy-ridden
countries it is necessary for the medical profession to learn to
diagnose leprosy, especially in its early stages. The public must be
persuaded of the fact that there is no credit or advantage to be
gained from having even only a few hundred or few thousand
lepers hidden away in the midst of the community.
In Johannesburg a prominent political man had a leper for
about ten years as his gardener; the boy was repeatedly diagnosed
as a syphilitic. The mixed case of leprosy figured in Abraham’s
chapter on the disease in Allbutt and Rolleston’s System of
Medicine, page 682, toured South Africa with an operatic company,
was laid up in Johannesburg with an acute leprotic attack, was
visited and treated by a medical man who failed to recognise the
malady. Numerous similar instances could be quoted, all showing
that, for a system of segregation to succeed, it needs above all the
intelligent co-operation of the general medical public. Without it
all efforts are of little use.
There can be no doubt that segregation in asylums, if properly
carried out, is in every case an expensive matter. No doubt it is
more expensive still, in the long run, if adopted in an incomplete
fashion, because the sum total eventually employed will have failed
to bring about a corresponding diminution of leprosy.
The Union of South Africa is at the present moment spending
quite £100,000 a year on its lepers (according to the estimates for
1913-14, £97,800), and is segregating certainly not more than half
the number of lepers which are believed to exist in the territories of
the Union. Under the circumstances the question arises whether
these stupendous efforts and large sum of money would not be better
employed in stamping out diseases which are more infective, such
as typhoid or syphilis, and show a better chance of responding to
treatment.
Unfortunately we are dealing with a subject which is but rarely,
if ever, approached without a certain preconceived view and
sentiment. The general public, on the one hand, considers leprosy
to be extremely contagious, shuns and has an absolutely fanatic
terror and horror of the disease. As a contrast, the patients them-
7 $
selves and their families cannot as a rule be persuaded to take the
simplest measures to prevent an immediate spread of the scourge.
In face of this dilemma nothing but segregation seems to meet the
requirements of both parties.
An alternative to isolation in asylums is home segregation,
which, however, as a rule, fails to have the desired effect if not
aided by constant inspection, which may become so irksome that
many prefer to take refuge in asylums. In South Africa it has not
been a success, though in Norway it has been possible to introduce
the system on a large scale.
When dealing with numerous lepers, such as are to be found
among native populations, a system of segregation in asylums does
not seem to be advisable on the score of efficiency, expense, and
practicability. In these instances isolation in settlements with
natural boundaries (a river, steep hills, on islands, etc.) appears to
be by far the best solution. Wives can then accompany their
husbands and the usual amemties of kraal-life (cattle, agriculture)
need not be dispensed with. A central hospital for the infirm and
blind, a dispensary for dressings and ambulatory medical
attendance, a home for untainted children, should be provided, but
the inhabitants of the settlement, whether lepers or healthy, would
have to work for themselves; the only restriction placed on them is
that they are not allowed to leave a certain district or province.
Amongst natives generally, especially in South Africa, pure
maculo-anaesthetic leprosy is by far the commoner type of the
disease. Many of these anaesthetic cases are by no means incapable
of work; in fact, graduated exercise and manual labour is distinctly
beneficial to their general good health. In asylums, strapping
young natives, with a simple contracture or a few maculae, slide
into a life of sloth, mope about the wards and yards, and if
re-infection can take place, certainly expose themselves to its danger
to an unnecessarily increased extent and jeopardise accordingly
their slender but possible chance of recovery.
Asylums for lepers cannot be done away with; they are equally
necessary, if not more so, than hospitals for the mentally afflicted.
Where feasible, they should be supplemented by settlements or
leper colonies, and in exceptional and carefully selected cases by
home segregation.
77
By these means there is every reason to believe that leprosy
would soon become extinct in any country, however a firm and
extensive hold it may have taken, however unsuccessful previous
half-hearted attempts of prevention may have been, however long
the scourge may have existed in the past.
XII. CONCLUSIONS
Though numerous questions are yet unanswered in the pathology
of leprosy, and many points are still under discussion, it cannot be
denied that the researches of the last ten or fifteen years have
opened up further fields of fruitful investigation, and helped to
elucidate several obscure problems.
It is true that the results of some experimental observations are
so diametrically opposed that it appears hardly possible that both
can be correct, at least not in their entirety. This is no new feature
in the study of diseases whose investigation presents more than
ordinary difficulties. Malaria is a good example. First of all
bacteria were isolated time after tinie from patients, and it was
sturdily maintained they were the etiological factor of the disease.
Then the protozoa in the erythrocytes were stated to be artefacts by
men eminent in various branches of medicine. Later on the
plasmodia were found in marsh-water and so forth. Finally, the
present conclusive standpoint was reached, but the path to truth
was strewn with the thorns of controversy and deeply indented with
the pit-falls of erroneous deductions.
This comparison explains the present position of leprosy
research. The exercise of sound and reasoned scientific criticism
ought to enable us to unravel, in course of time, the delicate and
elusive thread of positive fact from the tangled skein of mistaken
interpretations of essentially correct, but irrelevant observations.
The main conclusions which can be deduced from a perspective
review of recent experimental study of leprosy appear to be
concentrated in the following main points :
Cultivation of Hansen's ‘ bacillus'
Any micro-organism isolated from lepers which claims to
correspond to the acid-fast rod seen in lesions should be identified
78
by injection into animals and the following production of the well-
known histological features of lepromas.
The culture of Kedrowsky and those similar to this type are
the only ones to fulfil this essential postulate.
Clinical features of leprosy.
In addition to the nodular, maculo-anaesthetic, and mixed types,
a further variety is recognisable, wuth thick raised patches, which
may be more or less confluent and circinate, containing numerous
giant cells, and very scanty acid-fast rods.
Histological appearances of typical leprotic lesions.
In dermal nodules and corresponding lepromas of inner organs,
there occur numerous matted masses of acid-fast rods, intra-cellularly
and extra-cellularly situated, very slight tissue reaction and scanty
giant cells.
The lesions of inner organs which show caseation, necrosis or
numerous giant cells of the Langhans type, appear in some cases to
be due to tuberculous complications. Guinea-pig inoculations only
can conclusively decide the nature of such appearances.
Maculae show the features of slight chronic irritation due to the
action of minute doses of bacterial toxins. This, in addition to the
external resemblance between maculae and the erythema produced
by injecting the water-soluble contents of various acid-fast cultures
into lepers, seems to postulate that leprotic maculae may be the
result of a local reaction similar to the one resulting in von Pirquet’s
and similar tests for tuberculosis.
Rat leprosy.
This spontaneous disease presents numerous points of resem¬
blance to leprosy, and may yet be found to be etiologically related
to the human malady.
Its experimental transmission in rodents only causes analogous
but not identical effects to the spontaneous disease.
Transmission to animals.
The negative results of numerous experimental inoculations of
human beings, and the relative low infectivity of leprosy, should
prepare us to face very numerous unsuccessful results in dealing
79
with this question. It is only by injecting a large series of animals
in various ways, and then observing them for prolonged periods,
that deposits of acid-fast ‘bacilli’ can be produced in the inner
organs which have the essential histological features of the leproma.
Serology.
Wasserman’s test is incapable of distinguishing in every case
syphilis from leprosy. The same applies to Eitner’s test. Noguchi’s
luetin test is negative in lepers whose disease is not complicated by
syphilis.
Leprotic sera often show spontaneous absorption of complement.
The serological distinction of leprosy and tuberculosis is not
possible in every case. Tuberculine tests may be present in lepers
showing no clinical symptoms of tuberculosis.
Agglutination is so far of little value owing to self-clumping of
acid-fast bacteria and low titer of leper sera.
Contagiousness.
The communicability of leprosy by direct and indirect contact,
especially under defective hygienic conditions, has been established
by numerous repeated and independent clinical observations.
Treatment.
Leprosy, especially the macular variety, is subject to spontaneous
remissions and self-cures.
Chaulmugra oil is indicated for nodular cases; cultural extract
for macular lepers (as far as the present experience goes).
Prevention .
Registration and sanitary inspection of lepers’ habitations;
segregation in asylums, settlements, or colonies; early separation
of children from leprous parents; betterment of hygienic conditions
where leprosy has spread extensively.
8o
Fig.
Fig.
Fig.
Fig.
Fig- 5
Fig. 6
EXPLANATION OF PLATES I-VI
Plate I
Filamentary interlacing, branching, non-acid-fast germ cultivated
from the nodule of a leper on horse-serum-nutrose-agar. Some of
the original acid-fast rods from the leprous lesion are still to be
seen and the acid-labile bacteria have grouped themselves in a
peculiar fashion round the 1 globi.’ This culture was injected into
a rat, acquired acid-fast properties, and could be then regained in
pure culture as an acid-fast ‘ bacillus.’ Stain: Ziehl-Neelsen.
Magnification, i.ooo diameters.
Diphtheroid, pleomorphic, filamentary and bacillary germ, isolated
from the nodules of a leper. It is considered to be a further develop¬
mental stage of the previous micro-organism. These bacteria are
slightly acid-resisting, and regained acid-fast properties after injec¬
tion into animals. Third sub-culture on placental-juice-glycerine-
agar. A slight multiplication of the original acid-fast rods may
have taken place. Stain: Ziehl-Neelsen. Magnification, 1,300
diameters.
Acid-fast stage of the bacteria of leprosy which are commonly met
with in tissues. (‘ Hansen’s bacillus,’ Mycobacterium leprae,)
Pure culture with which all identification experiments have been
carried out. Two months’ culture at 37°C. on fish-juice-glycerine-
agar. From a micro-photograph. Stain : Ziehl-Neelsen. Magnifica¬
tion, 500 diameters.
Section of omental gland from a spontaneously leprous rat, found
in Ipswich by Drs. Petrie and Macalister. The similarity to the
lesions seen in human leprosy is very striking. Stain : Ziehl-
Neelsen. Magnification, 500 diameters.
Nodules produced in the kidney of a rabbit by the intravenous
injection of a culture of ‘ Moeller’s smegma bacillus.’ This lesion
simulates leprosy in one detail only : the presence of massed acid-
fast micro-organisms. Stain: Ziehl-Neelsen. Magnification, 500
diameters.
Section from the skin of a leprous rat, showing the enormous
quantities of micro-organisms which cause the lesions. The thick,
purple masses consist almost entirely of ‘ bacilli ’; the blue spots
are the nuclei of the connective tissue.
From a colour-photograph taken by Sanger-Sheppard’s method.
Stain : Gram. Magnification, 400 diameters.
[Plates I-IV have already been published by the author in the S. Afr . Med,
Record (1913), XI, pp. 201-222.]
Annals Prop. Med, & Parasitol ., Vol. IX
PLATE I
6 .
82
Plate II
Fig. 7. Lesions produced in the liver of a rabbit after intravenous injection
of the acid-fast culture of 4 Hansen’s bacillus.’ Similar appearances
can be brought about by the intravenous injection of cultures of
human and avian tuberculosis, and under circumstances by the
inoculation of leprosy * virus.’ Stain : Ziehl-Neelsen. Magnifica¬
tion, 500 diameters.
Fig. 8. Lung of a mouse which has been injected intraperitoneally with
Kedrowsky’s strain of leprosy. The .enormous quantity of 4 bacilli ’
present has caused no appreciable tissue reaction of a tuberculous
nature. Stain : Ziehl-Neelsen. Magnification, 150 diameters.
Fig. 9. Spleen of a monkey injected intraperitoneally with Kedrowsky’s
strain of leprosy. Also in this instance the numerous bacteria have
not brought about a tissue reaction of a tuberculous nature. Stain :
Ziehl-Neelsen. Magnification, 35 diameters.
Fig. 10. Omental gland of a rabbit, which received an intravenous injection
of Kedrowsky’s strain of leprosy. The section is teeming with
acid-fast ‘ bacilli ’ which have not caused the death of the cells they
have invaded, because the nuclei are still capable of staining, and
have not appreciably altered in morphology. Stain : Ziehl-Neelsen.
Magnification, 25 diameters.
Fig. 11. Spleen of a mouse which had been injected intraperitoneally with
an emulsion of Kedrowsky’s strain of leprosy. The red patches are
groups of cells which are crammed with acid-fast rods. Similar
lesions are sometimes seen in the spleen of lepers. Stain : Ziehl-
Neelsen. Magnification, 25 diameters.
Fig. 12. Portion of the mouse spleen shown in fig. 11, but under increased
magnification. The ‘ bacilli ’ are seen to have nearly completely
filled the protoplasm of the cells without inducing necrosis or
caseation. No giant cells of the Langhans type are to be detected.
These are the points which on the whole distinguish this lesion
from one of 4 tubercular ’ nature, i.e., caused by Koch’s 4 bacillus.’
Stain : Ziehl-Neelsen. Magnification, 230 diameters.
Annals Trap. Med. & Parasitol., Vol. IX
PLATE II
.
9 .
12 . ir
8 4
Plate III
Fig. 1.3. Localised lesion produced in the testes of a rat by the injection of
human leper ‘ virus.’ Stain : Ziehl-Neelsen. Magnification, 500
diameters.
Fig. 14. Leprous lesions in the spleen of a rat six months after injection of
a ground-up inguinal gland from the preceding animal. Drawing
made from three different portions of the slide. The peculiar
hyaline increase in size of protoplasm of the cells invaded by the
4 bacilli’ is specially noticeable. Stain : Ziehl-Neelsen. Magnifica¬
tion. 500 diameters.
Fig. 15. Section through the skin of a rat suffering from spontaneous rat
leprosy. The acid-fast 4 bacilli 9 of the disease have congregated
round the shaft of a hair-follicle. (According to Borel may have
been deposited there by Demodex .) Stain : Ziehl-Neelsen. Mag¬
nification, 35 diameters.
Fig. 16. Small, solitary deposit of 4 bacilli ’ in the omentum of a rat, one
month after intraperitoneal injection with ground-up 4 virus ’ from
a spontaneously infected rat. The micro-organisms are markedly
granular, yet juice of tha spleen of this animal produced a pure
culture of the germ, the only one achieved out of very numerous
attempts. Stain : Ziehl-Neelsen. Magnification, 700 diameters.
Fig. 17. Thorax gland of a white rat injected intraperitoneally with an
artificial culture of rat leprosy. The lesion is identical with that
produced by the injection of 4 virus.’ Stain: Ziehl-Neelsen.
Magnification, 500 diameters.
Fig. 18. Small deposit of acid-fast micro-organisms detected in a nodule of
the abdomen of a mouse injected with an emulsion of a leprosy
culture which had been heated for an hour at 6o°C. Showing
that, killed or alive, leprosy micro-organisms are capable of
producing similar lesions. Stain : Ziehl-Neelsen. Magnification,
300 diameters.
Annals Trop. Med. & Parasitol., Vol. IX
PLATE III
«
i?
tf
4
*
$
#
» 4
#
• I
V
86
Plate IV
Fig. 19. Filamentary, interlacing, branching, partially acid-fast and acid-
resisting germ isolated from the pleura of a man, by Birt and
Leishman. It will be seen that it shows marked clubbed fragments
(so-called involution forms), and also some which are definitely
‘ bacillary’ in appearance. The resistance to the bleaching pro¬
perties of mineral acids varies also considerably. From a pure
culture on glucose-agar. Stain: Ziehl-Neelsen. Magnification,
800 diameters.
Fig. 20. Acid-fast bacteria from the water-tap at the Lister Institute. Here
also marked pleomorphism and variable acid-fastness is noticeable,
but in this case the culture is not known to be pure, because the
micro-organism was not isolated on artificial media. Similar acid-
fast rods have been found in the scraping from a water-tap in the
Public Health Laboratory at Cape Town and in Stockholm;
also in mule’s dung at Robben Island. These observations show
the ubiquity of acid-fast rods. Stain : Ziehl-Neelsen. Magnifica¬
tion. 600 diameters.
Fig. 21. Section marked * Experimental Leprosy, Monkey,’ sent by Professor
Duval, and considered by this author to be diagnosable as leprosy.
Stain : Haematoxylin-Eosin. Magnification, 60 diameters.
Fig. 22. True branching in tubercle-bacillus from sputum of consumptive.
From a drawing by Mr. W. D. Severn. Stain: Ziehl-Neelsen.
Magnification, 2,250 diameters.
Fig. 23. Pure culture of Duval’s strain of leprosy. Its coccoid appearance
shows it to be morphologically very similar to the ubiquitous group
of saprophytic acid-fast micro-organisms, therefore quite different
from ‘ Hansen’s bacillus.’ From a micro-photograph. Stain:
Ziehl-Neelsen. Magnification, 1,100 diameters.
Fig. 24. Mononuclear cells from the peritoneal cavity of a guinea-pig, forty-
eight hours after injection with a chromogenic strain of an acid-fast
micro-organism. (Duval’s 4 lepra.’) Stain : Ziehl-Neelsen.
Magnification, 1,800 diameters.
88
Plate V
Fig. i. Front and back view of an off-coloured Cape-boy, with serpiginous
or lupoid leprosy. The presence of nerve-lesions and similar
appearances on single other cases of leprosy, allow the inference
that the disease is due to an infection through Hansen’s ‘ bacillus,’
though acid-fast micro-organisms are very scarce indeed in the
skin-eruption.
Fig. 2. Leprous-patch of face, apparently taking its origin from the eye, in
a Fingo native. The borders are more raised and the surface
rougher than is usually the case in leprosy. Similar patches are
present on the loins of this case. No anaesthesia or paraesthesia
detectable.
Fig. 3. Arm of nodular leper 24 hours after intradermal injection of 10
minims of cultural extract. There is a copper-coloured areola
round the site of injection, in the centre of which a small sore has
developed.
Fig. 4. Arm of an arrested anaesthetic leper, 24 hours after intradermal
injection of 10 minims of cultural extract. Numerous blisters;
intense local reaction, rise of temperature to ioi° F., malaise, pains.
All symptoms passed within three days.
Annals Trop. Med. & ParasitolV61. IX
yo
F>g- 5
Fig. 6
Plate VI
Diagrams of case before treatment (12.9.12). On face, infiltrated
and thickened patches, red in colour. On legs, erythematous
patches, reddish in colour. Over feet, purplish flush.
Diagrams of case after treatment (6.5.14). There are faintly
pinkish-red flat discolorations on the face. There is a flat brown
macula on right upper arm, below the shoulder. The other yellow
discolorations are very difficult to chart, as many of the edges are
not well defined. There is a faintly light purple flush on the limbs,
and the insides of the hands are a darker shade of the same.
jhifi [)
H A n
'. m ^ n m
.rJU va'vj f w
9 1
THE METABOLISM OF WHITE RACES
LIVING IN THE TROPICS
I.—THE PROTEIN METABOLISM
BY
W. J. YOUNG
FROM THE BIOCHEMICAL LABORATORY, THE AUSTRALIAN INSTITUTE OF TROPICAL MEDICINE,
TOWNSVILLE, N. QUEENSLAND.
(Received for publication 2 November, 1914)
The effect of high temperature and a moist atmosphere upon
the metabolism of the human body has been studied by workers in
temperate climates, the conditions being artificially produced in a
specially constructed chamber, and also by a few investigators who
carried out experiments upon both acclimatised whites and natives
living in the Tropics. In almost every case attention has been
given to the total exchanges of the body, with the object of
ascertaining whether the altered conditions bring about the intro¬
duction of any new factor into the mechanisms by which the body
regulates its thermal equilibrium.
The most important investigations done in the Tropics were
those of Eijkman in 1893, who determined the calorific value of
the everyday food of a number of white men living in Java, and
found that the average did not differ from that usually accepted
as a standard in Europe. Examination of the food of Malays
also showed that the heat value per kilogram of body weight was
not materially different from that of the white men. In these
researches balance sheets were drawn up between the total nitrogen
taken in and that excreted, but no investigations were made into
the separate nitrogenous products of protein metabolism.
A number of analyses of the nitrogenous constituents of the
urine were made by McCay (1908) in India, but were confined to
an examination of the urines of Bengalis, a people who conform
to a diet with much lower protein content than Europeans.
9 2
In the course of a study of the effect of residence in tropical
countries upon a working white race, it was thought that some
information might be obtained by a fairly complete examination
of the partition of the nitrogen and sulphur excreted in the urine.
The significance of the different excretory substances in urine is but
very imperfectly understood, and the effect of any departure from
normal conditions is of interest from the possibility of throwing
light upon the processes which constitute the nitrogenous
metabolism of the body.
The principal product by which proteins give up their nitrogen
in the body is urea, and the bulk of this substance appearing in
the urine represents the ammonia produced by deaminisation of
nitrogenous foodstuffs. It tells us nothing as to what has
happened to the remainder of the molecule from which it has been
derived.
Certain substances, however, appear to be produced by the
metabolism of the tissues, the endogenous metabolism of the body.
Thus, the creatinine is almost entirely derived from this source,
since the quantity of creatinine excreted on a low protein diet is
practically the same as when the diet is rich in nitrogen (Folin).
Only a small fraction is due to creatine taken in with the food.
Leathes (1907) found that during fever, when the tissue metabolism
is known to be increased, the quantity of creatinine in the urine was
also increased, and recent experiments on rabbits by Meyers and
Volovic (1913) have confirmed this observation, and have shown
that the output of creatinine is increased whether the pyrexia be
caused by infection or by confining the animal in a hot atmosphere.
The total quantity of sulphur contained in the urine normally
runs parallel to nitrogen, varying with the quantity and nature of
the protein catabolised. The bulk of the sulphur is excreted in the
fully oxidized form as inorganic sulphates, together with a small
quantity in combination with indol and skatol as ethereal sulphates,
these products being the result of bacterial decomposition in the
intestine. A small quantity of sulphur is excreted in an unoxidized
form, termed neutral sulphur, which in all probability consists
mainly of cystine and thiocyanates. The output of neutral sulphur
in normal individuals was found by Folin to be independent of the
quantity of protein in the diet, and for this reason he considered
93
this portion of the sulphur as derived, like the creatinine, from
endogenous sources.
From the experiments on creatinine mentioned above, it seemed
possible that tissue metabolism might take place to a greater extent
in a tropical climate, and it was of special interest to find whether
the quantity of the substances derived from this source was very
different from the averages in urines of residents in temperate
climates.
Fairly complete analyses were made of the daily urine of four
male subjects ranging in age from 25 to 36 years, and who had
resided in Tropical Queensland from one to four years. The
experiments extended in most cases over a week, the total urine
being collected and each twenty four hours’ sample analysed. The
subjects were living an ordinary laboratory life, were allowed to
choose their own diet, as the aim was to make the observations
under the conditions of every day life. Moreover, most investi¬
gators are agreed that the knowledge of the diet being controlled
is not without influence on the quantity of food taken, whilst if a
fixed diet be partaken, the continued sameness affects the appetite.
As a preliminary, therefore, the diets were uncontrolled as regards
quantity and character, excepting in one case (protocol IV) where
the subject was taking a milk and bread diet of known nitrogen
content.
In addition to these four experiments, determinations have been
made at different times of the total nitrogen and the creatinine in
the daily urine of five subjects, and in two cases comparisons have
been made of the quantities of these substances excreted during the
hot rainy season and the cooler dry season.
METHODS
The total nitrogen was determined by Kjeldahl, and urea by
Folin’s (1912) potassium acetate method, uric acid by the Folin-
Schafer method and the purin bases were estimated after precipi¬
tating the uric acid as recommended by Kennaway (1909).
For the estimation of creatinine Folin’s well-known method was
employed, but some modification was required on account of the
high temperature of the laboratory. Although numerous papers
94
have been published on the conditions under which this determina¬
tion should be carried out, such as the quantities of reacting fluids,
the time during which the solutions are allowed to react, the
temperature and the extent of dilution before the readings are
made, all such conditions have been determined for room tempera¬
tures about I5-20°C. In this laboratory, during the hot season,
the air temperature during the day is rarely below 27° C. and
generally ranges from 30 to 35 0 , whilst that of the water supply
is usually about 30°. The conditions laid down by other workers
were, therefore, not applicable, and had to be readjusted to meet
the requirements. For this purpose a solution containing
10 mgms. of creatinine in the form of the picrate (mpt. 205-206°)
in 20 c.c. was employed, and the depth of colour produced by the
addition of alkali and picric acid compared with a 01N solution
of potassium bichromate in the Dubosq colorimeter in the usual
manner. Experiments were also carried out with urine. It is only
necessary here to give the conditions established, without going
into the experimental details. It was found that at temperatures
between 27° and 35° C., 10 mgms. of creatinine gave the maximum
coloration within four minutes, whilst the colour faded if the
reacting liquids were allowed to stand for longer than six minutes.
The best quantities of the reagents were 15 c.c. picric acid
(saturated solution) and 5 c.c. of 10 % sodium hydroxide, the whole
being diluted to 500 c.c. as recommended by Folin. Within limits
the total volume of the reacting fluids did not influence the colour,
thus 5 c.c. of urine diluted either to 10 or 20 c.c. gave the same
reading. The colour of the diluted mixture was found to fade
fairly quickly, the solutions were therefore compared with the
standard immediately after the final dilution was made. Most of
the urines examined were very concentrated, and 5 c.c. usually
contained enough creatinine to give a reading on the colorimeter
between 7 and 9, the standard being set at 8. If, however, the
reading came beyond these limits the estimation was always
repeated with a larger or smaller quantity of the urine. The usual
cr 1 N potassium bichromate solution was employed as a standard,
and was carefully checked by a solution of pure creatinine picrate.
Of the other constituents the total sulphur was estimated by
Benedict’s method, the inorganic, ethereal and neutral sulphates by
95
Folin’s methods, the phosphoric acid by titration with uranium
acetate, and the chlorine by Volhard’s method. The gravity was
determined by Mohr’s balance.
The full details of the analyses are given in the protocols at the
end of the paper.
VOLUME, SPECIFIC GRAVITY AND TOTAL NITROGEN
Determinations of these have been made on the twenty-four
hours’ urine at different times, each experiment lasting from four
to ten days so as to obtain a fair average. Most of these were done
in the hot season of the year (November to April) when the
atmospheric humidity is high, but in two cases comparisons were
made in the cooler season (May to October).
The full details are given in the protocols, whilst the averages
for each set of experiments are collected in Table I, the number in
Table i. —Volume, Specific Gravity and Total Nitrogen of the Urine
24 Hours Urine
No.
Subject
Age
■
Weight
kilos
Protocol
Season
Nitrogen
in Grams
VoL
cc.s.
Spec,
gra v.
Total
Per kilo
of body
weight
i
A
3 2
66-8
1
hot
986
1,023
11*69
0*175
2
A
33
66-8
2
hot
772
1,027
11*58
0*173
3
A
33
66-8
3
hot
1,070
1,024 *
14*33
0*214
4
B
2 5
6o*6
5
hot
922
1,027
12*41
0*205
5
D
3 °
63-1
8
hot
7%2
1,026
10*83
0*172
6
D
l
3 o
62-8
7
cool
*,437
*,017
12*11
0*193
7
E
36
48-2
11
hot
621
1,023
op
00
r*->
OO
6
8
E i
35
48-4
9
cool
>>037
1,021
11*69
2*242
9
E
36
48*2
1°
cool
1 , 37 2
1,012
10*52
0*218
IO
C
2 7
62*2
! 6 |
cool
1
i ,3 67
i
1,014
11*02
°* I 77
Average for both seasons, 5 persons =
1,070
1,021
ii *49
0.194
Average for hot season, 4 persons =
817
1,025
Ii*i 5
0
So
00
Eijkman's average for 19 Europeans in Java =
1,442
1,017
13*04
0*200
Pfluger and Bohland for Europeans
—
—
12*67
0*194
Bleibtreu and Bohland for Europeans
—
—
H *93
0*233
9 6
the fifth column referring to the protocol containing the full data.
For comparison are also given the figures obtained by Eijkman for
Europeans living in Java, as well as those for Europe found by
Pfliiger and Bohland and Bleibtreu and Bohland. The volume of
urine varies from day to day, but the average figures show that
during the hot humid weather, when any exertion is accompanied
by profuse sweating, the urines are comparatively small in volume
and of high gravity. The volume of liquid taken in was generally
between 3 and 4 litres a day.
Nos. 5, 6, 7, 8 and 9 show the effect of the season on the
excretions of the same individuals. The total nitrogen also shows
daily variations, but the averages, 8*8 to 14*3 grams, are somewhat
lower than that given in the physiological text-books as the average
for Europe, 14 to 18 grams. When the nitrogen is considered in
relation to the body weight it does not differ appreciably from that
found in Europe. In the cases examined the daily excretion of
nitrogen was greater in the cool season than in the hot. Part of
this difference is no doubt due to the nitrogen lost in the sweat
during the hot weather. Eijkman (1893) found that as much as
076 to 1*36 grams of nitrogen per day were excreted cutaneously
bv Malays, whilst Benedict (1906) outside the tropics, found that
during rest the nitrogen lost in this way was about 0*071 grams per
day, but during severe muscular labour as much as 0*13 to 0*22
grams per hour.
During the hot part of the year a nitrogen-balance experiment
was carried out on subject A. The numbers, Table 2, have not
Table 2. —Nitrogen balance on fixed diet.
Date
Nitrogen in
food
N
[trogev Excreted
Balance
Urine
Faeces
Total
4 - 3*14
26*58
17*01
2-99
20*00
+ 6*58
5 * 3*14
26*58
18*87
1*50
20*37
4* 6*21
6.3.14
26*58
19*24
3*52
22*76
+ 3-82
7 * 3*14
26*58
20*32
1*82
2214
+ 4-44
Total .
+ 21*05
97
been included in the general averages, since the diet, which was
restricted to milk and bread and butter, was of much higher
nitrogen content than the usual diet. The subject increased in
weight by 1*3 kilos during the four days, and complained of feeling
uncomfortably full after the meals. The numbers show a consider¬
able nitrogen retention amounting to 21 grams, a small part of
which may be accounted for by the cutaneous excretion.
UREA
The excretion of urea varies with the total nitrogen.
The average daily outputs in the different subjects varied from
2698 grams in A to 1479 grams in E, or 87*9% to 78*4% of the
total nitrogen excreted respectively. The quantity of urea
generally quoted as the average for Europeans on a mixed diet is
30 grams per diem; the quantities found were always less than this
amount, but when considered in relation to the total nitrogen
appear quite normal.
AMMONIA
The ammonia present in the urines was quite normal in amount,
the averages varying from 0*50 grams to 0*64 grams, or 4*8%. and
3*6% of the total nitrogen respectively. The average output for
all four subjects being 0 57 grams.
CREATININE AND CREATINE
Creatine was only present in any of the urines on one occasion,
and then only in a very small amount (012 grams nitrogen).
A number of determinations of the creatinine have been made
both in the hot weather and in the cooler season. Table 3 gives
the averages over the whole periods, given in detail in the
protocols, each figure representing the average output per day for
a period of several days. These averages vary between 0*47 grams
and 071 grams of nitrogen per day, representing between 3*2 and
6*1 % of the total nitrogen. In normal individuals in temperate
climates the daily excretion is, according to Folin (1905), from
3’5 to 4 5 %, whilst van Hoogenhuyze and Verploegh (1905) found
values of 4 to 6%. Leathes observed that although during fever
there was an increase in the actual amount of creatinine excreted,
9 8
there was at the same time increased nitrogen, so that the
percentage of the nitrogen which appeared as creatinine was actually
diminished.
If the creatinine in the urine be considered in relation to the
body weight, column 8, it is seen that the quantity excreted varies
in the different individuals from 8^52 mgrms. to io'62 mgrms.
per kilo of body weight, values which agree with the figures given
by Folin, namely, 7 to 11 mgms. per kilo of body weight.
Table 3. —The Excretion of Creatinine
No.
Subject
Season
Protocol
Total N.
grms.
Creatinine Nitrogen
grms.
per cent,
of total
N.
per kilo of
body wt.
mgrms.
1
A
hot
1
1 i-6q
'
o-6i
54
9’ 1 3
2
A
hot
* !
11*58
0-71
6-i
10-62
3
A
hot
3
*433 j
0-62
44
9-28
4 #
A
hot
4
1886
0-63
3*2
943
5
B
hot
5
1241
0-64
5 *
10-56
6
D
hot
8
10-83
o -57
5*2
9*°3
7
D
cool
7
I2*II
0*58
4-8
9-23
8
E
hot
11
8-83
o *49
5.6
10-16
9
E
cool
9
j II-69
0-47
4 ‘*
971
10
C
cool
6
| I 1 -02
0-53
4*9
8-52
• No. 4 on special milk and bread diet -» 26*6 grms. N.
As in all these experiments, with the exception of No. 4, the
diets contained creatine, the quantities of creatinine excreted are
probably slightly higher than that which represents tissue meta¬
bolism alone.
The figures, therefore, so far as they go, supply no evidence of
a greater creatinine output in a tropical climate, nor do they show
any marked seasonal variation.
URIC ACID
The average quantity of uric acid varied from 0*42 to 0*56
grams per day, which is quite a normal figure for a mixed diet.
99
PURIN BASES
The considerable variations observed in the daily excretion of
the purin bases are to be attributed to the tea and coffee of a mixed
diet, since the greater part of these substances in the urine arise
from this source. This is confirmed by the fact that with A on
mixed diet (protocol III) these substances varied from 58 mgrms.
to 92 mgrms. per day, whilst on a bread and milk diet (protocol IV,
the quantity was much less and more constant, and varied during
the next three days only from 21 to 28 mgrms.
PHOSPHORIC ACID
The phosphoric acid varied very considerably, the average on
mixed diet being between 1*56 and 2*61 grams per day. The
nitrogen to P a 0 6 ratio usually accepted for male Europeans is
5 or 6 : 1; in every case but one the average ratio found lay between
5*5 and 5*9, the exception showed a ratio of 71.
CHLORIDES
The quantity of chlorides excreted was too variable in amount
to allow of any conclusions.
SULPHUR
The total sulphur when compared with the nitrogen gave ratios
N : S of 14 to 15, this is somewhat higher than the average given
in the text-books on Physiology, viz., 12*75, but Cathcart and
Green (1913) during a number of determinations of this ratio, when
different proteins were superimposed on a fixed diet, found in some
cases numbers as high as this.
The quantities of neutral sulphur excreted were rather high,
representing on a full protein diet from ii*8 to 20*8 % of the total
sulphur, and averaging from 0*250 grams to 0*455 grams per day
as S 0 3 . The average quantity on a full diet is, according to Folin,
about 0*25 grms. per day, or about 5 % of the total sulphur. On a
low protein diet he found no lessening in the amount excreted, the
quantity then representing 25 % of the total sulphur. The
inorganic sulphates and the ethereal sulphates were quite normal in
amount.
100
CONCLUSION
The number of subjects upon which the experiments were carried
out is, of course, too few to draw any definite conclusions, but as
far as they go they exhibit no marked variations from the averages
obtained in temperate climates. In every case the neutral sulphur
was high, but whether this is due to an increased tissue metabolism,
or not, only further experiment will show.
REFERENCES
Benedict (1906). Joum. Biol. Chem., I, 263.
Cathcart Sc Green (1913). Biochem. Joum., VII, 1.
Eijeman (1893). Virchow’* Arch., CXXXI, 170.
- (1893). Virchow’* Arch., CXXXIII, 105.
Folin (1905). Amer. Joum. Physiol., XIII, 66, 117.
- (1912). Joum. Biol. Chem., XI, 507.
Kennaway (1909). Joum. Physiol., XXXIX, 296.
Leathes (1907). Joum. PhytioL, XXXV, 205.
McCay (1908). Scient. Mem. Gov. India, No. 34.
Meyers Sc Volovic (1913). Joum. Biol. Chem., XIV, 489.
Van Hoogenhuyee Sc Verploegh (1905). Zeit. Physiol. Chem., XLVI, 415.
PROTOCOLS
I
Subject A.—33 years ; weight «* 66*8 kilos ; four years in Tropics
Date
* 9*3
Volume
CC.S.
Specific
gravity
Total
nitrogen
grms.
Creatinine
grms.
As
nitrogen
grms.
per cent,
of total
nitrogen
Mar. 19
'>487
1,015
14-20
1*65
o*61
4 * 3 2
„ zo ...
i, 4*5
1,013
10*25
**47
o *55
5 ‘ 3 2
„ 21 ...
1,030
1,028
* 5 * 3 1
i* 5 *
0*56
3-67
• 9 22 -
775
1,027
9**7
i *59
0-59
6-44
,, 2 3 -
690
1,026
9-52
**55
OO
U">
6
6*05
,, H -
*75
1,025
12*07
1*56
0*58
4*80
91 25 ...
745
1,026
9-61
1*89
0*70
7 * 3 *
„ 26 ...
880
1,027
12*12
i *99
o *74
6*io
„ 27 ...
970 ;
1,022
12*93
*•52
o *57
4*37
Averages ...
986
*,023
11*69
1*64
o*61
5-38
II
Date
* 9*3
Volume
CC.S.
Specific
gravity
Total
nitrogen
grms.
Creatinine
grms.
As
nitrogen
grms.
per cent,
of total
nitrogen
Dec
. 16
730
1,029
iz -43
2*24
00
6
6*70
,,
17 ...
675
1,028
1**31
*•93
i
072
6*34
,,
18 ...
895
1,024
I2*38
2*00
o *74
6*oo
,,
19 ...
755
*,027
9*60
i-6 7
0*62
6*46
,,
20
*75
1,026
12*04
I * 9 2
071
5 * 9 2
,,
21
820
1,028
ii *37
2*02
°*75
6*6o
,,
22
720
1,030
11*26
I78
o*66
5*86
,,
23 ...
705
1,028
12*24
173
0*64
5* 2 5
Averages ...
772
1,027-5
i.- 5 8
I *91
o*71
6 14
102
Averages] 87*9 3*6 4-4 i*o | 0-5 | 2*7 76*14 7*68 17*60
Bread and Milk Diet
104
Averages 81-5 4*1 5-1 1-5 0-5 7-3 76-94 8-47 14-58
io5
VI
Subject C.—27 year* ; weight — 62*2 kilos ; 2} years in Tropics.
Volume
CCS.
Specific
gravity
Total
nitrogen
gnns.
Creatinine
Date
*9*3
Grms.
As nitrogen
Percentage
of Total N.
July
30 ...
'.540
1,011
10 86
1*44
°*53
494
»,
31 ...
1,040
1,020
13*31
1 65
o*6i
461
Aug.
1 ...
1,170
1,0*4
10*37
1*40
0*52
504
,,
2 ...
1,480
*,014
11 3 *
**45
0-54
476
3 ...
1,100
i,o *7
10*00
*‘47
°*55
5*48
M
4 •••
'>*55
1,015
10*84
1 *46
0 54
5 02
»,
5 -
■> 43 °
1,014
10*25
1-44
053
5*22
6 ...
1,625
1,010
10*42
no
0*48
4 * 6 5
7 -
1,665
1,011
11*87
* *36 1
0*50
4.27
Averages ...
'>367
*,014
1 1*02
*•44
o*53
489
VII
Subject D.— 30 year* ; weight — 62*8 kilos ; 1-1J yean in Tropics.
Date, 1913
Volume
CCS.
Specific
gravity
Total
nitrogen
grms.
Creatinine
Grms.
As nitrogen
grms.
percentage
of Total N.
July 30 ...
2,340
1,011
1372
i*8o
0*67
487
„ 31 ...
97 °
1,024
1215
1*40
0*52
4-^7
Aug. 1 ...
1,540
1,018
12*28
i*66
0*62
5*02
„ 2 ...
*, 73 o
1,012
12*26
1*85
0*69
5*60
3 -
970
1,024
11*85
1*48
o *55
465
,, 4 •••
1,190
1,019
13*86
i *59
o *59
4 * 4 *
„ 5 •••
*, 3 *o
1,015
12*10
*•45
o *54
4*45
„ 6 ...
*, 43 o
1.015
11*51
1 *53
o *57
4*95
„ 7 -
1,950
1,012
97I
1-29 1
-
0*48
4*92
„ 8 ...
940
1,020
11-62
,. 5 o
0*56
479
Averages ...
*,437
*,o *7
12*1 I
**55
0*58
4*79
VIII
Mixed Diet; 63*1 kilos
12.4.14 84*4 3*2 5-3 i*4 o-8 4*9 66-07 *3* i8 *°* 6 5
107
IX
Subject E.—36 years*, 48*4 kilos; 1 to 1} years in Tropics
Date,
> 9*3
Volume
CCS.
Specific
gravity
Total
nitrogen
grms.
Creatinine
Grms.
As nitrogen
grms.
Percentage
of Total N.
July
6 ...
790
1,022
9*°4
1*28
0*48
5*3
»»
7 •••
9>5
1,023
10-47
1*32
0*49
47
8 ...
970
1,022
1179
1*28
00
rf
b
3-9
9
1,185
1.016
10*19
1*22
o *45
4‘5
10 ...
1,118
1,018
11*36
1*22
o *45
4 *o
11 ...
i,5>o
1,016
11*50
>*34
0*50
4*3
-
12 ...
895
1,024
12*60
1*28
0*48
3*8
13 ...
850
1 026
13*08
1*20
o *45
3*4
14 ...
*> 37 °
1,015
*4-30
1*21
o *45
3 ’*
15 ...
770
1,026
12*54
1 - 3 *
o *49
3*9
Averages...
>i°37
1,021
11*69
I *27
0*47
41
X
E. Weight = 48*2 kilos.
Date, 1914
Volume ccs.
Specific gravity
Total nitrogen
grms.
June 25 .
1,540
1,011
9*97
» 26 .
1,150
*j°>3
io*86
» 2 7 .v
1,460
1,012
io *77
„ 28.
'.340
>3013
10*50
Averages
*> 37 *
1,012
10*52
Mixed Diet
64-17
109
THE SPECIES OF PARAGONIMUS AND
THEIR DIFFERENTIATION*
BY
HENRY B. WARD
AND
EDWIN F. HIRSCH
(Received for publication 23 September , 1914)
Plates VII-XI
CONTENTS
Introduction .
page
. 109
Historical Summary.
. no
Material .
. "5
Structure or Paragonimus ...
. 116
General Form .
. 116
Cuticular Spines.
.. 118
The Alimentary System ...
. 123
The Excretory System ...
. 128
The Reproductive System
. l2 9
General Discussion .
. 146
The Genus Paragonimus ...
. 146
Cuticular Spines.
. 148
References .
. * 5 *
Explanation or Plates
. *54
INTRODUCTION
In 1895 the senior author published the first account of the
occurrence of a mammalian lung fluke on the North American
Continent. On various occasions since then he has contributed to
the knowledge of these forms. In 1910 the junior author found
some of these parasites in a host which was indisputably a native
of Wisconsin, and became much interested in the problems
connected with the discovery, so that when in the following year he
engaged in graduate study at the University of Illinois this topic
was naturally selected for his work. The present paper represents
the results of that work and of the contemporaneous and subsequent
Contributions from the Zoological Laboratory of the University of Illinois, No. 34.
no
studies of the senior author. Unfortunately, even after very
considerable delay, it has proved impossible to obtain all of the
material from distant regions essential for comparison and deter¬
mination of all the species which are involved in the literature of
the subject. But we feel clear that the results, even if imperfect
still in certain details, should be laid before scientific workers in
this field without further delay that they may be tested by those
more favourably situated to secure material in quantities.
HISTORICAL SUMMARY
The earliest record of a lung fluke in mammals was published
by Kerbert in 1878. During an autopsy held in the Zoological
Gardens at Amsterdam, Holland, in September, 1877, parasites
were found in the lungs of a tiger, and were sent by the Director,
Westerman, to Kerbert, who determined them to be undescribed
trematodes and named the species Distoma westermanii. Three
years later Kerbert received specimens, through Bolau, from the
lungs of a tiger that had died in the Zoological Gardens at
Hamburg, Germany. Upon this material, which proved to be
identical with the former, he completed an extensive morphological
study (Kerbert, 1881) of the species.
The first information concerning the occurrence of such a
parasite in man came from Manson, who in a letter to Cobbold,
published with comments by the latter, described (Manson, 1880)
the discovery of distome eggs in the sputum of a Chinaman suffering
from hemoptysis. At the same time he mentioned a fluke found in
another patient, a Portuguese, by Dr. Ringer. The specimen was
sent to Cobbold, who in the note just cited named it Distoma
ringeri. Although the description is scanty, the records leave no
doubt as to the identity of the species. It should be noted that
this is the first published record of this human parasite, ante¬
dating by nearly two months that of Baelz, to which priority is often
accorded.
The latter record came thus almost simultaneously, but from
Japan. As early as 1878 Baelz had noted there conditions, similar
to tuberculosis and regularly attributed to it, which he regarded as
resulting from some parasitic infection. He sent the sputum to
Ill
Leuckart for examination and described the bodies in it as psoro-
sperms, but before receiving Leuckart’s diagnosis, came to the
conclusion that they were in reality eggs of some fluke. This
opinion was first published by Manson (1882), who stated that after
having examined some of Baelz* gregarine material he had found
bodies identical with the ova of Distoma ringeri. In the following
year Baelz (1883), confirmed in his view of the nature of the material
by the report of Leuckart, published a description of the worm, which
he re-named Distoma pulmonale . According to Stiles and Hassall
(1900, p. 567), certain Japanese investigators in the interval had
named the species Distoma pulmonis . Both this name and that of
Baelz are ante-dated by the name of Cobbold, which is apparently
the first designation used for the human parasite. There is no
need to review here the important clinical and pathological data
submitted by Manson, Baelz, and later writers. The wide and
abundant distribution of the parasite in Japan, Formosa and Korea,
its occurrence among all ages and classes of society, and the high
ratio of the infected in certain districts are well-known and firmly-
established facts. The Asiatic lung distome is one of the most
important human parasites.
The next important step in the history of this form was taken
by Leuckart, who in conjunction with his student Nakahama made
a careful study of specimens of the fluke which Baelz had sent him
and of specimens of Distoma westermanii from Kerbert. Although
he noted points of disagreement, these were interpreted as
unimportant minor differences, and the specific identity of the two
forms was definitely asserted. We do not find that this identity
has been questioned since then, except once (Ward, 1908). Stiles
and Hassall (1900, p. 561) voice the general opinion in stating that
the human parasite ‘ though originally supposed to represent a new
species is now generally admitted to be identical with Kerbert's
form from the tiger.*
Regarding the genus, however, several investigators commented
independently on the impossibility of retaining this form in the old
group Distoma ; and finally in 1899 Braun made it the representa¬
tive of a new genus to which he gave the name Paragonimus The
first careful and extended description of the genus came from
Looss (1899), who unaware of Braun's publication gave the form
112
another name that, because of its slightly later appearance, must
be relegated to synonymy. However, the description given by
Looss (1899) still ranks as the most accurate and complete avail¬
able. Even at that time Looss recognized only a single species,
to which he gave the name Paragonimus westermanii *
Except for its sporadic extra-limital occurrence in hosts that
were known to have come from the regions where the parasites are
endemic, the species was not recorded from the western continent
until one of us (Ward, 1894) published a description of a similar
form obtained from a cat in Michigan. This paper called attention
to the differences between these worms and the description given by
Kerbert and Leuckart for Distoma westermanii , but assigned them
tentatively to the same species. It also noted the possibility that
the host had been brought as a pet from the East, and that hence
the parasite was not en'demic. Grave doubt was thrown on this
view through the discovery by Kellicott of worms in the lung of a
dog from Ohio, which were sent to Ward and pronounced by him
identical with those he reported from the cat.
All possibility that the parasite had been found only in hosts
introduced from abroad disappeared when Stiles and Hassall
(1900) recorded the abundant discovery of a similar parasite from
hogs slaughtered at Cincinnati, Ohio. Their paper gives not only
a very careful account of these specimens but a detailed comparison
with other records and the best critical summary of the literature
on these lung flukes which has been published to date. While
recognizing the possibility that their worms ‘ may represent a
distinct variety at least,’ Stiles and Hassall 1 feel compelled to
continue for the present to look upon the American form as identical
with the Asiatic.'
Since then the lung fluke has been reported in numerous cases
from North America. In the human host it was first diagnosed by
Mackenzie, in 1904, from the lung of a Japanese fisherman on the
Columbia river. Then Fehleisen and Cooper (1910) reported a
case in a Japanese worker in California fruit orchards who had
come to the country some six years before. Microscopic examina¬
tion of the sputum showed in every field two or three eggs of the
distome. No details of size or structure are given, but the case was
• Looss wrote P . westermanni which is evidently a typographical error.
n 3
of long standing and had been diagnosed in Japan as lung fluke.
Thus all the evidence goes to show that it was not contracted in the
United States. In the same year Null (1910) recorded its
occurrence in a Korean at Seattle, Washington.
From other hosts there are several cases on record. Null stated
casually that it occurs in dogs and cats from the Oriental quarters
of San Francisco, but gave no further data regarding the parasites.
Nickerson (1911) recorded the occurrence of P . kellicotti in the lung
of a cat from the grounds of the University of Minnesota at
Minneapolis. Only three specimens were found.
The junior author of this paper (Hirsch) observed several cases
of marked significance in Wisconsin. As early as 1907 he noticed
a peculiar cough in a pet cat, which became more seriously affected
and was chloroformed a year or more later. These worms were
found in the lungs. A kitten born to the first cat was affected in
the same way and found to be infected with the lung fluke when
only a little more than a year old. In addition to these cases, of
which the full case history, written by Hirsch, was published by
Hanson (1911, p. 112), the former has also found the lung fluke in
two other cases hitherto unpublished. These cases are another
kitten of the same litter and a fourth cat, unrelated and living some
miles distant from the first three. It is positively known that the
first three were born in that locality and had never been away from
it, hence the endemicity of the parasite can only be questioned on
the ground that possibly the means of infection—a fish carrying the
encysted young distome for instance—was brought in from a
distance. There is no probability that the two young cats were
infected directly from the mother cat, but in all likelihood through
the consumption of the same food. It seems improbable that all
three could have been infected at the same date, and certainly the
fourth cat did not acquire its parasites from the same food. These
cases are the first in which the presence of the lung fluke was
diagnosed in the living animal and the diagnosis confirmed by the
demonstration of the ova in the sputum. They are also the first
on this continent in which the place of birth and infection was
positively determined for the host.
The range of the human lung fluke was further extended by the
work of Musgrave (1907). He studied seventeen cases with eight
autopsies in the Philippine Islands, and gave a careful morpho¬
logical description of the parasite. His data will be utilised later
in our paper.
One of the seventeen hosts was a Chinaman, two were Japanese,
and the other fourteen were native Filipinos, so that the species is
undoubtedly endemic in the Islands. Various later papers contain
casual references to the occurrence of this species in the Islands,
but do not give data of value for our purposes. Garrison and
Leynes (1909) studied the development of the ova of Paragotiimus
in the Philippines. They do not give a precise description of the
ova, but apparently these were obtained from cases among those
recorded by Musgrave (1907) which have already been considered.
The work of Garrison and Leynes, which deals with the experi¬
mental development of Paragotiimus ova under varying conditions
of temperature, light, salinity and desiccation, has bearings of great
importance on the dissemination of the disease and the infection
of man, but does not throw any light on the problem of species and
their differentiation.
In a paper published several years ago, the senior author (Ward,
I 9°3) commented on the great discrepancies between the measure¬
ments recorded by various investigators for the eggs of Paragotiimus
westermanii , and gave an outline sketch representing these differ¬
ences in a graphic manner. He reached the conclusion that all
records could not be accepted as correct unless more than one species
was concerned. The measurements given by Yamagiva, which were
made from ova in sections of the brain and lungs of man, have
since been shown to belong, in all probability, as Ward stated later,
to the Japanese blood fluke, Schistosoma japonicum , and are
consequently eliminated from the present discussion. In a later
paper (Ward, 1908) these records were discussed at greater length
in the light of further evidence, and the conclusion was reached that
the American form originally identified as Paragotiimus westermanii
is undoubtedly a distinct though closely-related species, and to it
Ward gave the name of Paragonimus kellicotti. He also indicated
the probable specific independence of the Japanese form on which
further work was then being done. This form, which will be
fully discussed in the descriptive section of this paper, must bear
the name of Paragonimus ringeri (Cobbold, 1880).
MATERIAL
The junior author made a detailed and careful study of
Paragonimus kellicotti on new material obtained from the pig at
the Cincinnati (Ohio) abattoirs. The specimens were taken alive
from the lungs, and after shaking in normal salt solution according
to the method of Looss, were preserved with great care. The
technique involved nothing unusual, and gave good results
throughout. Total preparations and serial sections of the
specimens originally obtained by Ward from the cat in Michigan,
by Kellicott from the dog in Ohio, and by Hirsch from the cat in
Wisconsin, were compared item by item with this new American
material.
Several specimens of the Japanese form from man were also
available for comparative study through the courtesy of Dr. S.
Uchida, of Tokyo, and eggs from the Seattle case were kindly
sent us by Dr. Null. This form has been described in detail several
times, notably by Katsurada (1900) and Kubo (1912). Despite
this fact, the structural features are not even yet well known, and
in most respects these descriptions are couched in such general
language that we cannot determine from the paper more than
generic features regarding certain organs. The last paper especially
(Kubo, 1912) falls short of what might be wished. In spite of a
rich supply of fresh material he gives very little more information
on the structure than the work of Katsurada a dozen years earlier.
The figures are distinctly unsatisfactory, being in some cases vague
and almost illegible, and in others highly diagrammatic. They
are certainly inferior to those given by Katsurada.
Our work was supplemented by comparison with three co-types
of Distoma westermanii kindly placed at our disposal by Professor
Kerbert and now in the Ward collection. While this supply was
adequate for the determination of the most important features in
the anatomy, as will appear in the following pages, it did not
suffice for a complete study of the structure, and a detailed
comparison of this form with those from man in Japan, and from
cat, dog, and hog in North America, must be left to some future
student who has at his command a larger supply of material.
Despite persistent effort, it has been impossible to secure for
study and comparison any material from the Philippine Islands,
ii6
so that the status of that form could not be tested by the method
worked out on the other material. Although only a single paper
has been written on the anatomy of this form, that one is so
carefully worked out that it furnished very definite material for
comparison with the work on other species. In spite of this, there
are several points on which we would fain have had precise infor¬
mation concerning the structure of some organ that was not treated
in extenso in the text of that paper.
STRUCTURE OF PARAQONiMUS
General Form
Paragonimus kellicotti has a somewhat elongated form,
elliptical in dorsal aspect. The dorsum is strongly arched, the
highest point being somewhat anterior to the middle portion of
the body, while the ventral surface is slightly flattened. The
anterior end rounds off gradually, but the posterior extremity is
attenuated, and sharply curved. The range in size is given in
comparison with Paragonimus ringeri and Paragonimus westermanii
in the following table: —
Species
Host
Length in mm.
Width in mm.
Thickness in
mm.
Authority
Max.
Min.
Av.
Max.
Min.
Av.
Max.
Min.
Av.
P. kellicotti
Hog
I 1*0
8-5
9-8
3*5
3 *o
3 ‘i
3*2
2*2
2*6
Hirsch
P. kellicotti
Cat
1 5*5
11-2
136
77
4-8
—
—
—
—
Ward
P. kellicotti
Hog
i4-o
3 *o
—
4 -o
2*0
—
—
—
—
Stiles
P. kellicotti
Dog
20*0
i 5 *o
—
—
—
—
—
Kellicott
P. zvestermanii
Tiger
9-0
7 *o
—
6*o
4 -o
—
4 -o
2*0
—
Kerbert
P. ringeri ...
Man
10*0
8-o
—
6-o
V°
—
—
—
—
Leuckart
P. ringeri ...
Man
lyo
7 **
9-6
7*5
5 '°
5 *°
4 -o
3*5
37
Katsurada
P. ringeri ...
Man
'7
75
io-i
6
4*5
5*8
5-5
3*5
5
Kubo
11 7
From these figures it may be noted that there is considerable
variation in the size of the different individuals. Taken as a
composite picture, however, a description of the American form
emphasizes its much elongated, relatively slender structure, which
is in contrast with Paragonimus ringeri and Paragonimus
westermanii> both of which are oval-shaped, thicker, and broader
parasites.
The most recent and extensive study of fresh material for
Paragonimus ringeri has been made by Kubo (1912). He has listed
measurements of thirty-six specimens from the lungs of the dog in
Japan and of eleven specimens from the human lung in the same
region. There seems to be no contrasted difference between
specimens from the two hosts. The entire series averages in
length 10*08 mm.,* in width 5*8 mm., and in thickness 5 mm. The
smallest he found measured 3 mm. in length and the largest 12 mm.
Worms with a length of 5 mm. and over are sexually mature,
having some eggs in the uterus. He has not included these
extremely small and presumably young specimens in his averages,
so that the latter represent fairly well full-grown adults. Yet even
with that, the omission of five conspicuously small specimens listed
in his table will raise the size averages appreciably.
,On the other hand, when one examines a group of these parasites
with the eye it is not difficult to see a general difference between
the species in type of form. Yet at the same time it is clear that
neither the size nor the form gives a safe basis for distinguishing
the species. There is no doubt that some of the conflicting views
in regard to size are due to the measurement of specimens at
different ages and stages of growth.
At the anterior extremity is found the oral sucker directed
towards the ventral surface at an angle of about 45 0 , while the
acetabulum lies on the same surface in the median line slightly
anterior to the centre of the body. In ten specimens these suckers
average respectively 075 mm. and 0*83 mm. in diameter. The two
suckers of Paragonimus westermanii are of about equal size,
078 mm. in diameter, according to Leuckart, while those of
Paragonimus ringeri are unequal, and smaller, the oral sucker
Not io-8 mm. as given by Kubo in the text.
being 0*53 mm. in diameter and the acetabulum 0*6 mm., or at
most 075 mm. according to Leuckart.
Kubo says that the oral sucker of Paragonimus ringeri in a
medium-sized worm (10 mm. long) measures 075 mm. in diameter,
and that the ventral sucker in a similar specimen reaches 0 8 mm.,
being thus slightly larger than the oral sucker. These measure¬
ments differ from those of Leuckart and agree closely with those
we report for Paragonimus kellicotti , but here again the character
varies so much with the size and age of the specimen measured that
no dependence can be placed upon it in determining the species.
The genital pore in Paragonimus kellicotti lies just behind the
acetabulum, medial or a little to the right or to the left. The
position of the genital pore as given by Stiles for other species is
the same. The cuticula covering the entire body is relatively
thick, and armed with spines (PI. X, fig. 18). The thickness of
this structure, however, is not uniform over the entire body.
Around the suckers it may be 0005 mm. thick, while over the
posterior extremity it may be 0*048 mm. There is often also a
distinct difference in the appearance of this layer. Sometimes it
seems perfectly homogeneous, but in other places it appears as if
made up of two distinct layers, an outer more dense and less
refractive, and an inner vertically striated, rather clear, or highly
refractive with numerous coarse granules. The cuticula shows no
cellular structure, but is sharply marked from the underlying
muscles by a distinct basement membrane.
Cuticular Spines
Since a preliminary study of the spines revealed characteristics
that appeared to be of great value for the differentiation of species,
these structures were subjected to a most precise examination, with
results that we believe justify the labour. It should be recalled
that Kerbert, Leuckart, and Stiles differ in their statements
concerning the distribution of these structures and the place where
the largest are to be found, but so far as we can ascertain no one
has noted differences in the form of individual spines or in their
grouping. An examination of the three worms with reference to
these specific features gave pictures interesting both for their
resemblances and for their striking differences. The conditions in
the parasite from the lung of the pig are given first.
The spines of Paragonimus kellicotti lie in irregular circular
rows over the body and are set firmly in the cuticula with their
free ends directed posteriad. They extend entirely through the
cuticula, and sometimes into the body musculature beneath (PI. X,
fig. 18). Structurally, the spines are thin chisel-shaped scales,
usually several times as long as wide. The surface directed away from
the body is slightly convex, while that directed towards the body
is correspondingly concave. The free end is rather deeply serrated
into a number of very sharp teeth. These certainly aid the
parasite materially in effecting its movements, and in maintaining
its position in the tissues of the host. The basal end of the spines
is broader and thicker (0*005 to 0*010 mm.) than the tip, often
appearing cleft so as to show in cross-section groups of closely
aggregated oval or irregularly shaped chitinous bodies.
There is a distinct difference in the distribution and size of the
spines (PI. VIII, figs. 5-11) in various parts of the body. The suckers
are entirely devoid of these structures, while the cuticula closely
surrounding shows the transition stages between the non-spined and
the spined condition. This holds true especially for the region
surrounding the acetabulum. Here the body for a small distance
around the sucker may be entirely free from spines. When they
appear, they are few in number but very sharp and even decidedly
hooked (fig. 10). Around the oral sucker the spines (fig. 9) are
short with broad bases and sharp tips, a condition which is soon
replaced by larger spines with broad serrated tips. The spines are
set relatively more thickly over the anterior half of the body
(fig. 5) than over the posterior (fig. 7). The cuticula on the dorsal
surface just behind the oral sucker, and on the ventral surface
between the suckers (fig. 8) and just behind the acetabulum, is
especially well armed. The spines over the dorsal surface show
little variation except in number and in length. On the ventral
surface the greatest variation occurs around the suckers. This
divergence, however, is not one from the characteristic type, but
rather is a gradual diminution in size. The spines on this surface
of the body are perhaps shorter and broader than over the dorsum.
120
The results of an extensive series of measurements of the spines
in Paragonimus kellicotti have been brought together in the form
of a table or summary. This shows the exact size of individual
spines in various regions of the body, their distance apart, and
their variation in those cases where marked differences occur
(Table A).
Table A.—Spine Measurements of Paragonimus kellicotti
Distance apart 1
Length
Base
Tip]
Ventral Surface—
Near Oral Sucker.
0*005— Q, oo8 mm.
o*oio mm.
0*005 mm.
o*ooo mm.
Between Suckers.
0*013—0*026 mm.
0*046 mm.
0*013 mm *
0*013 mm.
Acetabular Region.
0*030 mm.—variable
J
0*010—0*020
o*ooi mm.
o*ooo mm.
Between Acetabulum and Anterior End
0*026 mm.
mm.
0*031 mm.
0*013 nim *
0*008 mm.
Between Acetabulum and Posterior End
0*013—mm.
0*030 mm *
0*023 mm.
0*021 mm.}
Posterior Extremity .
0*026—0*047 mm.
1
0*036 mm.
0*015 mm *
0*008 mm.
Dorsal Surface —
Anterior Extremity .
0*008—o*oi2 mm.
0*031 mm.
1
o-oio mm.
0*008 mm.
Anterior One-Fourth .
0*012—0*026 mm.
0*034 mm.
o*oio mm.
0*008 mm.
Middle of Body .
0*015— O‘oz6 mm.
0*044 mm *
o-oio mm.
0*008 mm.
Posterior One-Fourth .
0*026—0*036 mm.
0*040 mm.
o*oio mm.
- 0*008 mm.
Posterior Extremity .
o*oi2 -0*026 mm. *
0*036 mm.
o-oio mm.
0*008 mm.’
The spines of Paragonimus kellicotti are of one general type,
chisel-shaped, with distinctly serrated or saw-toothed free
extremities. There is, to be sure, a variation in their length, but
in general the characteristic features of this type of spine are
retained throughout. The study of these spines involved their
examination over the entire body of the parasite; in no place was
there any marked divergence in form, and nowhere did they occur
other than singly. While the substance of the comparison was
made upon material taken from the hog, further data were obtained
by examining a piece of the cuticula from a parasite removed from
the lungs of a cat. The shape of these spines, as well as their
121
arrangement, was the same (PI. VIII, fig. n), the only difference
noted was that in extreme cases they were somewhat longer
(C078 mm.), and their free ends more deeply serrated. Such
differences may easily be due to the age of the individual parasite.
Our material was not extensive enough to enable us to test the
question of a possible growth in the size of these spines during the
life of the adult parasite. But we think all will grant that this
extreme of variation in the cuticular spines is not great enough to
justify regarding them as separate and distinct from the type
found in the hog, much less to conceal their essential agreement
with that type. Their radical dissimilarity with the other types
will appear conspicuously when those types have been described.
The dimensions of the spines of Paragortimus westermanii, as
given by Kerbert, have elicited considerable comment, for he
reports that the largest spines were o‘oi8 mm. long by o - oo2 mm.
broad at the base, and the others were o'oio mm. long. He
describes the spines as lancet-shaped. Kerbert’s figure shows the
posterior end of the parasite and the surface for a short distance
around the oral sucker entirely devoid of spines. Upon
re-examining the co-type specimens of Paragonimus westermanii
in the Ward Collection, these statements were found to need some
emendation. The spines are indeed lancet-shaped, but are
considerably longer than Kerbert’s dimensions indicate. They are
fully as long as those in the cuticula of Paragonimus kellicotti,
being 0‘047 to 0^049 mm. long by 0 005 to 0 010 mm. broad at the
base. In addition, spines cover the entire body excepting the
suckers, as in Paragonimus kellicotti. Leuckart gives the measure¬
ments of the longest spines of Paragonimus ringeri as 0 06 mm.
with a base of 0 014 mm. broad. These dimensions approximate
the size condition revealed in Paragonimus kellicotti by our
investigations.
To judge from these data one would be forced to maintain that
there was no clear difference between the spines in the three species,
since all of the measurements fall well within the extremes we
found in Paragonimus kellicotti. And yet when one takes into
account the form of the individual spine and the arrangement of
the spines on the surface of the body there are evident and
distinctive differences of a striking character. The measurements
122
already given show that the spines of Paragonimus westermanii
and those of Paragonimus ringeri do not differ in size much, if at
all, from those of Paragonimus kellicotti, In the first two species,
however, the spines are markedly lancet-shaped, while in the last-
named they are, as already stated, broad and chisel-shaped. The
condition in Paragonimus ringeri is different. Over certain parts
of the body, especially on the ventral surface behind the
acetabulum, the spines are short with broad bases and tips. Here
they are set very closely together, forming at times almost
continuous circular rows. Along the side of the body the spines
are slender with sharp free ends.
While these differences in shape are very striking characteristics,
the spine arrangement is even more distinctive. The spines in
Paragonimus westermanii lie rather sparsely scattered in more or less
incomplete circular rows (PL VIII, fig. 13). Their free ends extend
very far beyond the cuticula, giving the parasite a thorny appearance.
In Paragonimus ringeri (PI. VIII, fig. 12) the general appearance
is very different, since the spines are characteristically arranged in
groups which are often massed together, and in certain parts of the
body are close enough to form almost continuous circular rows.
While the shape of the individual spines in Paragonimus ringeri is
subject to some variation, in fact offering a transition series
between Paragonimus westermanii and Paragonimus kellicotti , the
group arrangement affords a striking and constant means of
differentiation between the species.
In his description of the Philippine form, Musgrave (1907,
p. 32) discusses the distribution of the spines at length. He says
that on the ventral surface between the acetabulum and the oral
sucker they are ‘ almost entirely absent,* and that when they occur
in this region they are 'smaller than the others,* whereas they
increase in size on the lateral borders and reach a maximum on the
dorsum. This does not agree with conditions in the forms we have
studied and have described in the foregoing, as is shown distinctly
by a comparison of this statement with our figures. Musgrave
speaks of these spines as ‘scale-like,* a term difficult to compare
with our findings, and he gives no illustration of these structures.
From the evidence at hand we are unable to reach a final
conclusion, but it favours the separation rather than the amalgama-
123
tion of these forms with any known species. The decision of this
matter must await the study of material from the Philippine
Islands, which as yet we have been unable to secure.
In regard to the Japanese form, Kubo gives no data that can
be compared with our findings, but in one part of this description
he says that in larger and older worms the spines lose their sharp
points and become stumpy. This might suggest the interpretation
of the conditions we have described as possibly changes with
growth and use, dependent thus merely upon the age of the
specimens. This view is hardly tenable, since our illustrations are
taken from specimens of the same approximate size, and presumably
of the same age, if all were a single species. Furthermore, this
explanation does not touch the striking differences in the arrange¬
ment of the spines on the surface of the body.
The differences in form and arrangement of the cuticular spines
in the species we have studied may be summed up in tabular form
as follows:
P. westermanii
P. ringeri
P. kellicotti
Shape ...
Lancet-shaped
Chisel-shaped
Chisel-shaped
Very slender
Moderately heavy
Heavy
Distribution.
Sparsely, somewhat
irregularly, singly
Circular rows, in
groups
Circular rows, singly
The Alimentary System
The opening leading from the oral sucker into the pharynx in
Paragonimus kellicotti measures 0*050 to 0*075 mm - diameter.
It becomes somewhat larger toward the outer margin of the sucker,
and in sectional view appears wedge-shaped. It is continuous
proximally with a thin lamella that forms a small pocket, the pre¬
pharynx, which is located between the pharynx and the sucker and
serves to unite the two. No such region is mentioned by other
authors, and yet may be found in Paragonimus westermanii or
Paragonimus ringeri on careful examination.
The entrance into the pharynx is provided with four lip-shaped
124
projecting folds which effectively close the canal during contraction.
The two which lie laterally are larger than those bounding the tube
dorsally and ventrally, thus making the entrance into the pharynx
appear much like a vertical slit. The pharynx is spherical in
shape, about 0*4375 mm. in diameter. Its walls are composed of
heavy muscles, and the canal itself is limited to a narrow vertical
slit. The inner lining of the pharynx consists of a thin layer of
the cuticula, o*oi to 0*02 mm. thick. Towards its union with the
oesophagus the opening through the pharynx widens considerably.
The oesophagus is nearly circular in trans-section, and about
0*21 to 0 255 mm. long. The anterior part of the alimentary canal
is directed towards the dorsal wall, but not very acutely. After
the junction of the pharynx with the oesophagus this direction is
continued, and becomes a little more abrupt. At first the
oesophagus is about 0 08 mm. in diameter, but towards its
branching this dimension decreases slightly and then increases
again to 0*16 or even 0*175 mm. The thickness of the wall varies
in this short distance from about 0*012 mm. near the pharynx to
005 mm. somewhat nearer the bifurcation. This difference
necessarily causes the inner lumen to vary inversely. Such
relations as have been described are subject to variations no doubt
due to the contraction of these parts. Structurally the oesophagus
(PI. XI, fig. 23) is relatively simple, consisting of an inner homo¬
geneous layer, often thrown up in folds, and an outer well-
developed muscular layer. The latter is made up essentially of the
inner circular fibres, while the outer longitudinal fibres surround
the wall in relatively heavy parallel bands. This portion of the
alimentary canal is enveloped by well-developed glandular cells
whose ducts open into the oesophagus, in all probability furnishing
a salivary or digestive secretion.
„ The two lateral branches of the oesophagus are 0*175 to
0*255 mm * l° n g> an d are sharply differentiated from the intestinal
caeca by several distinct characters. The intestinal caeca
immediately attain a diameter of 0*315 mm., whereas the
oesophageal branches measure only 0*05 mm. in this dimension.
More generally speaking, there is in this region a sudden increase
in the diameter of the caeca to about four or five times that of the
oesophageal branches. In addition, the intestinal epithelium is
125
characterized by tall columnar cells (PI. X, fig. 17) which contrast
strikingly with the lining of the branches of the oesophagus.
The further course of the caeca is readily followed on the
reconstructions (PI. IX, figs. 14, 15, 16) made from a typical
specimen of Paragonimus kellicotli that was imbedded and
sectioned without having been subject to pressure or other
distortion. After the bifurcation of the oesophagus into the two
lateral branches, these ascend rather rapidly toward the outer and
upper margins of the body, and come to lie close under the
vitellaria, but not widely separated from each other. This
relation is apparent inasmuch as the body of Paragonimus
kellicotti in a natural condition is not broad or flattened but
attenuate and rounded. After its short transverse passage, each
of the intestinal caeca passes backward toward the posterior
extremity of the body, where they end blindly, one on each side,
quite close together. The termination of each canal takes place at
about the same level, although a slight variation occurs and
sometimes one terminates a little sooner than the other. In lateral
view the caeca show three large loops arching toward the dorsal
surface. Between these, secondary loops appear. The three
principal loops, while at first directed dorsally, ultimately curve
with the body wall, and when viewed from above are seen to be
turned also toward the median line. At these points the intestinal
caeca approach each other very closely; in fact, the space between
the most anteriorly located loops is very small. With this relation
in mind it is evident that the digestive system is more extended than
anticipated, and that each of the caeca if drawn out straight would
reach nearly or fully twice the length of the entire body. There
are also places where the intestine widens out considerably and
shows distinct enlargements. In these places the diameter may
become 0*68 mm., while towards the posterior extremity it is
reduced to 0*225 mm.
We are not sure how far it is possible to compare our
description of the alimentary system in Paragonimus kellicotti with
that of the other forms given by previous writers. We are not
unconscious that those descriptions are possibly less detailed rather
than actually different. What is certainly significant in explaining
some variations is that much in previous accounts is taken from a
126
study of specimens flattened under pressure. Such specimens must
be badly distorted, since this worm has a thick fleshy body and is
reduced by the pressure to a mere fraction of its normal thickness.
Nevertheless we feel that a comparison is worth while, and yields
some minor evidence that is significant and helpful in the solution
of our problem. On the whole it may be said that the anatomical
study of the alimentary canal in Paragonimus westermanii and
Paragonimus ringeri reveals some small but characteristic differences
from that of Paragonimus kellicotti . The pharynx is distinctly
smaller than in the latter. In Paragonimus westermanii it averages
0'5 mm. long by 0*3 mm. broad, according to Kerbert; it is, then,
not a spherical structure, as may be inferred both from these
dimensions and from Kerbert’s figure. The pharynx of Paragonimus
ringeri , according to Leuckart, is spherical and 0 3 mm. in
diameter, but Kubo givfcs it as elongate, measuring 0*4 mm. by
0*3 mm. The oesophagus is also shorter in both others than in
Paragonimus kellicotti , being in Paragonimus westermanii 0*14 mm.
long (Kerbert), and in Paragonimus ringeri 0*02 mm. long according
to Leuckart.
Kubo was the first to see that the regions of the two branches
immediately following the median oesophagus are identical
histologically with that canal, and should be counted a part of it
and not of the intestinal caeca which continue them but are so
different in structure. He gives a length of 0‘3 mm. for the
undivided region and 0*2 mm. for the lateral branches. He
comments on the sharp transition from the oesophageal to the
intestinal region, and ascribes to the latter a diameter two to three
times as great as we found in Paragonimus kellicotti . The peculiar
condition shown in both these forms in that the intestinal caeca do
not arise immediately from the end of the oesophagus, but some¬
what further distad from the branches of the median oesophageal
canal, is no doubt a generic character. It is well illustrated by the
figure of this region which Kubo gives.
Some features of the intestinal caeca constitute possible
differences. Kerbert says that in Paragonimus westermanii the
intestinal caeca pass posteriad from the bifurcation of the
oesophagus parallel with the surface of the body. They reach
nearly to the posterior extremity of the body, and then end
127
blindly. He gives the length of the caeca at about 8 mm., or
approximately the length of the whole body. This description is
certainly at least incomplete, for the caeca in Paragonimus
westermanii as he figures them do show some irregularities, and
perhaps this condition varies with the age of the parasite in all
species of this genus. But proceeding from the account of Kerbert,
one may say confidently that such a simple relation does not exist
in Paragonimus kellicotti or in Paragonimus ringeri. Leuckart
shows, both in his description and by his sketches, that the
intestinal caeca of Paragonimus ringeri are complicated by a
number of loops and turns just as they are in Paragonimus
kellicotti . Yet one point must be borne in mind, namely, that the
latter parasite is the more attenuate. In it the intestinal caeca
therefore do not lie very widely separated from each other, and
where the major loops arch through the body towards the median
dorsal line only a very small space intervenes.
One may ask if the formation of the loops is the same in both
species. According to Kubo the course of the intestinal caeca in
Paragonimus ringeri is entirely irregular and very varied. This
condition is shown in his figures, and may be a real difference
between that species and Paragonimus kellicotti . We hesitate to
accept this interpretation, since his figures are drawn from much
flattened preparations, and in such the general symmetry of the
caeca which we have described is so much modified that it can
rarely, if ever, be seen. It is possible that an examination of
undistorted specimens of Paragonimus ringeri will show some
regularity and balance in the course of the intestinal caeca. For
the present one must accept the statement that in Paragonimus
ringeri the course of the intestinal caeca is irregular and
asymmetrical.
These relations between the species are expressed synoptically
in the following table:
P. westermanii
P. ringeri
P. kellicotti
Size of Pharynx
o*3 x o*5 mm.
0*3 x 0*3 mm. or
0*4 mm.
0-44 x o*44 mm.
Length of Oesophagus
0*14 mm.
0*2 or 0*3 mm.
0*21 — 0*25 mm.
Character of Intestinal
Relatively simple,
Looped irregularly (?)
Looped symmetrically
Caeca
little longer than
body (?)
Twice length of
body (?)
Twice length of body
128
The Excretory System
The excretory system in Paragonimus kellicotti is relatively
simple. The excretory pore is of such size (0*05 mm. in diameter)
that under favourable conditions its position may be determined
with the unaided eye. It lies on the dorsal surface 0*2 to 0*225 mm.
from the posterior extremity, and opens into a small canal about
0 062 mm. in diameter and 0*225 mm. long. A cuticular layer lines
the canal, surrounding which is a well-developed muscular layer.
The canal passes directly forward, terminating abruptly approxi¬
mately midway between the dorsal and ventral margins of the large
excretory sinus which lies in the middle axis of the body. The
greatest dimension of this sinus is in a dorso-ventral direction, in
which also it approaches the body surface very closely, coming
within 0*15 mm. dorsally and 0*315 mm. ventrally behind the
acetabulum. It extends anteriorly to within 0 45 mm. of the
branching of the oesophagus, tapering to its termination here
gradually, and finally disappearing nearer the dorsal than the
ventral surface. As already stated, the sinus lies in the median
axis, but in the region of the uterus, which is located on the left
side somewhat anterior to the middle of the body; it shows a
distinct bending to the right. Its median position is again resumed
beyond the uterus. This bending is undoubtedly conditioned by
the increase in size of the uterus as it becomes filled with eggs.
The sinus has a distinct lining, as have also its main branches.
These branches are very numerous, though along the sides of the
sinus they are not conspicuous in the posterior region, but
anteriorly in front of the acetabulum it is easy to see a number that
are given off. These branches connect with the large stellate flame
cells that are distributed throughout the body. We could not
follow out the system to its ultimate details, but detected many
flame cells in sections.
The excretory system of Paragonimus westermanii consists also
of the prominent elongated central reservoir and numerous lateral
branches; in general a similar condition exists in Paragonimus
ringeri and, as we have shown, in Paragonimus kellicotti .
Considered more closely, however, there are here also certain minor
differences. According to Kerbert the large central reservoir in
Paragonimus westermanii is located in the posterior part of the
129
body, and opens to the exterior through a circular opening at the
posterior pole. Kerbert also does not mention the presence of a
short duct between the sinus proper and the excretory pore. This
may be a minor error in observation, or due to methods of technic.
But the appearance of the reservoir in Paragonimus westermanii is
in most cases, according to Kerbert, that of an elongate or pear-
shaped tube, or it may also be a spherical bladder. The latter
observation was made upon fresh material. The excretory bladder
in Paragonimus ringeri , according to Leuckart, is a very much
elongated narrow sinus with its greatest dimension in the dorso-
ventral direction. A short canal leads from the posterior extremity
of the sinus to the excretory pore. This opening has a diameter of
005 mm., and lies on the ventral surface. Our specimens of
Paragonimus ringeri are slightly distorted, but seem to indicate
this relation. In Paragonimus kellicotti the excretory pore appears
on the dorsal surface in about the same relation with the posterior
extremity.
Kubo locates the pore at the posterior end. He also describes
the canal system as originating from only two large main canals
which empty one on each side into the posterior portion of the
reservoir. Other authors have seen many such lateral canals
emptying into the central reservoir, and our findings in Paragonimus
kellicotti agree with them.
The other relations are expressed in synoptic form as follows :
P. westermanii
P. ringeri
P. kellicotti
Location of
Posterior pole
Ventral, near posterior
Dorsal, near posterior
Excretory pore
extremity, or at
posterior pole (?)
extremity
The Reproductive System
With the exception of the vitellaria,* the female reproductive
organs of Paragonimus kellicotti extend only a little beyond the
second quarter of the body. The genital pore lies about 0 078 to
0*104 mm. behind the acetabulum, usually a little to one side of
the median line. This is the opening of the genital cloaca. To
• In this discussion we have retained the classic names for the organs although, since the
appearance of Goldschmidt’s convincing demonstration, it can hardly be doubted that these
designations are inappropriate and incorrect.
130
the right, and near the dorsal wall, is the ovary, while on the left,
nearer the ventral surface, appears the highly coiled uterus. At
about the level of the ovary, but in the median line, lies the shell
gland, from which the proximal portion of the uterus emerges.
The genital cloaca is a short flask-shaped structure only 0*2 to
0’21 mm. long. It opens to the surface of the body through the
genital pore, and receives the terminal ducts of both the male and
the female reproductive systems. The wall of this structure
consists of an inner homogeneous or granular layer, and an outer
muscular wall which is composed essentially of circular fibres. The
terminal portion of the vas deferens narrows to a small tube, the
ductus ejaculatorius, which enters the genital cloaca at about the
middle of the base. The metraterm, or terminal portion of the
uterus, enters near the base, usually on the side opposite to the
acetabulum. Cirrus and cirrus pouch are absent.
Stiles records the genital pore as ‘median, right, or left, in
specimens from hogs/ but in our specimens variance from the
median location is slight, and where it is found, easily attributable
to a slight distortion of the body surface. What Kubo calls the
ductus communis genitalis evidently corresponds to what we have
designated the genital cloaca. In our specimens it is certainly a
distinct structure and not a common canal formed by the junction
of the male and female ducts. It is very much smaller relatively
than shown in his sketch, as is demonstrated by its length, which
he gives as 0'4 mm. in Paragonimus ringeri .
The male reproductive system of Paragonimus kellicotti
consists of two testes that occupy the third quarter of the body,
two vasa efferentia leading from them, and the single terminal
vas deferens. The vasa efferentia unite near the dorsal margin of
the excretory sinus to form the vas deferens.
The testes lie one on each side of the body, and occupy nearly
the entire space between the intestinal caeca and the excretory sinus
in the posterior region of the body. The central portion of each
testis is located approximately midway between the dorsal and
ventral body surfaces. Their symmetrical arrangement is slightly
disturbed, inasmuch as the right testis is usually a trifle posterior
to the left testis. In this particular there is some variation, for in
one specimen the right testis was found to lie anteriorly to the left.
This variation will be discussed later. The form of the organs is
noteworthy (Pl. VII, fig. 4, also PI. IX). Long slender lobes extend
from the upper margin of the central mass; these are usually two
in number. From the point of origin they arch upward and back¬
ward through the parenchymatous tissue. The terminal ends of
the lobes are greatly enlarged, and frequently sub-divided into
large rounded lobules. Other such lobes, three to four in number,
extend from the ventral surface of the central mass. These
connections are not as long as those given off from the dorsal
margin, but their terminal ends are much more prominently
enlarged, and almost always show two or more lobules heavier
than those on the dorsal projections of the organ.
The vasa efferentia, which are two in number, corresponding
one to each of the testes, have a diameter of 0 026 to 0*046 mm. at
their point of origin. They spring from the middle portion of the
testes at about the same dorso-ventral level, although the right one
is a little longer than the left. Each after its origin ascends
gradually, and at the upper margin of the excretory sinus they lie
parallel to each other. At this point the right one crosses to the
left side of the body, and after both descend somewhat, they unite
to form the vas deferens slightly to the left of the sinus at a level
just below the shell gland and vertically above the genital pore.
The vas deferens is at the start a relatively large duct, 0*062 to
o* 1 mm. in diameter. It drops in general ventrad, arching first
towards the anterior extremity. Keeping to the left side it
approaches the acetabulum partly surrounded by the coils of the
uterus. Then directing its course posteriad it circles close to the
acetabulum towards the ventral surface, finally terminating in the
genital cloaca. During its passage to the genital cloaca, the
vas deferens shows a number of characteristic features. Just at the
posterior margin of the acetabulum it suddenly narrows to a
small tube, which becomes even smaller as the genital cloaca is
approached. This portion of the vas deferens is heavily
musculatured, no doubt functioning as the ductus ejaculatorius.
Surrounding the vas deferens in the region where it suddenly
narrows is a mass of glandular cells (PI. XI, fig. 21). These
gradually disappear toward the genital cloaca and probably
constitute the prostate gland. The inner cuticular lining of the
132
genital cloaca is continued into the ejaculatory duct, but farther
on the nuclei of cells appear, although the lining retains its
granular structure. A muscular layer consisting of circular fibres
completes the wall of the vas deferens.
The testes of Paragonitnus westermanii lie near the dorsal side
of the body behind the transverse vitelline ducts (Kerbert). In
structure they show five to six lobes. The right testis lies close
behind the transverse vitelline ducts, while the left one is found
nearer the posterior end of the body. For this reason it is possible
to differentiate between an anterior right, and a posterior left testis
which are distinct in Kerbert’s figure. It must not be forgotten
that this was drawn from a much flattened specimen. The position
and relation of the testes is different in Paragonitnus ringeri
according to Leuckart. Here they lie, nearly symmetrically, well
towards the posterior extremity of the body. They are not
confined to the dorsal region, but occupy the greatest part of the
space between the intestinal caeca and the excretory sinus. In the
dorso-ventral dimension they have considerable extent. Comparing
the figures given by Kerbert and by Leuckart, one sees a clear
difference in form. The testes of Paragonimus westermanii are
dense and the lobes more regular in form, while those of
Paragonimus ringeri are diffuse and irregularly lobed.
Kubo compares the testes of Paragonimus ringeri to an out¬
spread hand, and says they consist of four or five long lobes
radiating from a common centre. Neither his description nor his
figure will fit conditions in Paragonimus kcllicotti as we have found
them, but they agree more nearly with Kerbert’s account of
Paragonimus westermanii . Here again it is hard to say how much
true conditions are modified by the distortion of flattened prepara¬
tions, but the testes in Paragonimus kellicotti are very much larger
and both the central mass and the more numerous lobes are larger
and heavier than the same structures as figured in the other species.
Kerbert records that the vasa efferentia in Paragonimus
westermanii pursue a dorsal course, arching over the transverse
vitelline ducts; after several loops they approach the ventral surface
and unite to form a common seminal vesicle which is continued
into a short ductus ejaculatorius. The vasa efferentia in
Paragonimus ringeri , according to Leuckart, pursue no such a
133
course, in fact neither one arches over the transverse vitelline
ducts, while the left one drops gradually to the ventral surface
without ascending dorsally. In addition, the vasa efferentia in
Paragonimus westermanii are more slender, being o*oi to o*oi6mm.
in diameter (Kerbert), while in Paragonimus ringert they are 0 045
to 0*1 mm. in diameter (Leuckart). We do not feel sure to what
extent these supposed differences, which are in fact rather minute,
depend upon trivial errors in observation or description or upon
different conditions of contraction in the body, and how far they
indicate real variations in structure between these closely-allied
species.
As already stated, the vasa efferentia in Paragonimus ringeri.
according to Leuckart, do not pursue a symmetrical course. It is
important to examine this further. The right tube rises gradually
toward the outer margin of the shell gland, crossing close under
the transverse vitelline ducts, then drops almost perpendicularly,
approaching at the same time the median line, and under the
ventral margin of the shell gland unites with the vas efferens of
the opposite side. This one pursues a much simpler course, for
it does not approach the dorsal surface, but is directed downward
toward the anterior extremity and median plane; it crosses the
margin of the excretory sinus relatively far forward, and continuing
its ventral and median direction is finally united with the other
into the common duct.
This portion of Leuckart’s description might be construed in
either of two ways: (1) that the union of the vasa efferentia takes
place ventrally to the excretory sinus, or (2) that it occurs dorsally
to the sinus. The figure in the text illustrating this point shows
the first relation, while the description might be understood as
indicating either. Among the specimens of Paragonimus ringeri
in the Ward collection there was one which had been broken just
behind the genital pore. The anterior portion of this parasite was
sectioned, and the relation of the vasa efferentia studied. In this
specimen the ducts united dorsally to the excretory sinus. The
vas deferens drops ventrally surrounded in part by the coils of the
uterus. That there is opportunity for certain variation in the
relation of the vasa efferentia, is readily understood from the fact
that, with continued growth of the parasite, the uterus becomes
! 34
engorged with ova, finally pressing nearby structures out of their
original relationships.
The vitellaria of Paragonintus kellicotti are very extensively
developed. Not only do they cover the parasite laterally, but also
extend over the dorsal surface of the body, meeting both anteriorly
and posteriorly, and leaving but a very narrow space in the median
line which becomes broader just in front of the transverse vitelline
ducts, finally to disappear entirely toward each extremity, although
around the oral sucker there is also a free space. The ventral
surface presents a relation very similar to the dorsal, except that
on this surface the vitellaria do not meet near the anterior sucker,
and correspondingly do not approach the median line so closely.
The product of the vitelline glands is gathered up by many small
ducts, which gradually unite to form two main trunks on each
side, one arising in the anterior region and the other in the posterior
region. These converge toward a point a short distance in front
of the middle of the body, and unite here to form the large dorsally
located transverse vitelline ducts.
This distribution of the vitellaria stands in partial contrast with
the condition in Paragonintus westermanii and Paragonintus ringeri.
In these species, as described, a considerable space near the median
dorsal and ventral line is not covered by vitellaria. In other words,
the vitellaria on the dorsal surface of Paragonintus kellicotti
approach more closely the median line than do those of
Paragonintus westermanii and Paragonimus ringeri , and in other
ways also appear to be more extensive in their development. The
relation on the ventral surface is very similar. The vitellaria of
Paragonimus kellicotti approach the median line more closely than
those of Paragonimus westermanii and Paragonimus ringeri , but
at the same time not so far as they do on the dorsal surface (PI. VII,
fig. i).
The vitelline reservoir in Paragonimus kellicotti is a pear-shaped
structure arising at the point of union of the transverse vitelline
ducts. These ducts become considerably narrower just before
terminating in the vitelline reservoir. At the point where they
unite the reservoir has its greatest width; dropping ventrally for a
short distance and at the same time becoming narrower, it directs
its course anteriad. Having reached a plane just below the shell
!3S
gland, it changes its course, and proceeds slightly upward until
about the level of the junction of Laurer’s canal and the oviduct.
Here it turns sharply to the right, and unites with the short canal
formed by the union of these two ducts (PI. X, fig. 20).
This condition closely typifies the relation in Paragotiimus
westermanii , but not that described by Leuckart and Kubo for
Paragotiimus ringeri. Here the two vitelline ducts unite to form
a single canal, which drops ventrally a short distance, and then
broadens out into a large flask-shaped reservoir 0 5 mm. long.
From the median margin of the anterior extremity of this reservoir,
and on the inner side of the unpaired canal, a small duct arises
and passes dorsally to unite with the duct formed by the junction
of the oviduct and Laurer’s canal.
The ovary in Paragotiimus kellicotti lies on the right side, close
to the dorsal wall of the body. Only a small portion extends
down far enough to lie alongside of the excretory sinus. The
transverse vitelline ducts bound this organ posteriorly (PI. IX,
figs. 15,16). In relative size the ovary is about as large as a testis,
but not so diffuse. It presents a more compact form since, even
though lobed, the lobes are heavy and do not extend so far from
the main body of the organ as do those of the testes.
The oviduct is a short tube, at first relatively wide (o'36 mm.).
It arises near the upper margin of the ovary, and from that portion
which lies toward the median line. It soon narrows down to a
very small tube, 0 018 mm. in diameter and about o'13 mm. long.
Just after its origin on the ovary, the wall of the oviduct becomes
heavily musculatured. This portion of the canal is the oocapt.
The oviduct rises slightly toward the dorsal surface, but drops
again towards the plane at which it left the ovary. Here it unites
with a small duct, which in fact is the so-called Laurels canal.
The wall of the oviduct consists of the cellular lining as is
described for the male reproductive system, and the outer circular
muscle layer (PI. X, fig. 19).
Little can be said of these organs in a comparative way. The
ovary lies in the same region of the body in all three species. We
did not find any constant differences in its form or in the structure
or relations of the oviduct in the different types.
Near the beginning of Laurer’s canal is an expansion measuring
0 057 mm. in diameter, and from this pocket there extends outward
to the right a blind pouch, or small seminal receptacle, about
o'195 mm. long and 0*052 mm. in diameter. Laurer*s canal makes
its way from this pocket in a sinuous course towards the dorsal
surface. While at the expanded region the canal is relatively
large, it soon narrows down to a very small duct. It proceeds in
a large loop directed posteriad, followed by another small loop in
the opposite direction to a short vertical stretch directly above the
shell gland which terminates on the dorsal surface of the body in
the region of the transverse vitelline ducts. The expansion and
the seminal receptacle, as well as the entire lower portion of
Laurer’s canal, swarm with spermatozoa. The oviduct and
Laurels canal unite to form a tube about 0*031 mm. in diameter.
This extends only 0*045 mm - before it receives the vitelline duct,
and then widens to form the ootype. The ootype discharges into
the proximal end of the uterus.
Stiles and Hassall (1900) have stated that a receptaculum
seminis is lacking in the pig lung fluke. Their statement is not
surprising in view of the real condition, which is shown at a glance
in the figure representing the shell gland complex (PI. X, fig. 20).
It is not possible, so far as we can determine, to detect such a
structure in total preparations, but sections through this region
demonstrate beyond question the existence of a true receptaculum
seminis in its normal location. It has the form of a blind pouch
opening into LaurePs canal near the inner end of the latter organ.
At this point the canal itself is much expanded, and the receptacle
can hardly be said to possess a neck or duct. Consequently the
cavity of the pouch is in constant and open communication with the
lumen of the canal, and spermatozoa circulate freely in the
common space. The receptacle is small, measuring at most
0*195 mm * length by 0 052 mm. in maximum width, so that its
extreme tip does not even reach the border of the shell gland.
The condition shown by the receptaculum seminis is of great
interest from the standpoint of comparative anatomy. As is well
known, there has been much discussion regarding the function and
meaning of this organ. Most students regard it as a structure which
is not of present functional value in any important way, at least
among the trematodes, and certainly its variable character and
*37
occasional complete absence are strong arguments in favour of such
a view. In some cases Laurer’s canal is reported to be lacking, and
in others the receptaculum has been said to be wanting, as in the
present instance. The actual presence of so insignificant a sac is
only a theoretical correction of that statement. It may properly be
designated a mere vestige of a structure about to disappear entirely.
It is even possible, of course, that individual variation between
different specimens is present to a sufficient extent to reduce it still
further than the condition represented in the figure. So far we
have found no evidence of its variation in size in Paragonintus
kellicotti .
According to Kerbert, the lower portion of Laurels canal in
Paragonintus westermanii is provided with a seminal receptacle,
which he figures somewhat larger than we find it in Paragonintus
kellicotti. Leuckart, however, doubts the accuracy of this observa¬
tion, since he did not find such a structure in Paragonintus ringeri.
A seminal receptacle certainly is present in Paragonintus kellicotti ,
and Kerbert not only records its presence in Paragonintus
ivestertnanii , but also gives its measurements. If this structure is
not present in Paragonintus ringeri , its absence may mark a
characteristic difference in structure in the latter species.
We are rather more inclined to believe that its presence can be
demonstrated, and that if the three species differ at all in
this respect, it will be found to be in the degree of development,
or perhaps one should say, of reduction, which this organ
manifests. Yet Kubo states that, despite zealous search, it was
not possible for him to detect such a structure, and such a definite
statement creates a strong presumption that the organ does not
exist in Paragonintus ringeri.
The shell gland as reported for Paragonintus ringeri by
Leuckart is a large organ, lying a little to the right of the median
dorsal region of the body. It is 0*5 mm. thick and about 1 mm.
long. This organ is also well developed in Paragonintus kellicotti.
It lies close to the dorsal wall, and is more or less oval in shape,
although somewhat irregular in outline, and measures 1 mm. long,
0 5 mm. thick, and 0 87 mm. broad. Kubo makes it slightly
smaller, viz., o‘8 by o*6mm. Kerbert records approximately the
same location in Paragonimus westermanii , although the shell gland
>38
is distinctly smaller, being 0’2 to 0*3 mm. long by 012 to 014mm.
broad. The shell gland surrounds the proximal portion of the
uterus, as well as the terminal portions of those ducts which go to
form this part of the female reproductive system.
The uterus in a fully-matured parasite is a condensed, closely
coiled tube. As the ova accumulate the uterus becomes widely
distended, so that the coil occupies nearly the entire lateral portion
of this region of the parasite. The walls of the uterus arc made
up of a relatively thin cellular layer, and a well-developed
muscular coat consisting of circular fibres. Towards its terminal
portion the uterus narrows down to form the metraterm. In this
region the walls become heavily musculatured (PI. XI, fig. 22).
The relation of the uterine coils in Paragonimus westermanii ,
as indicated by Kerbert’s sketch, is apparently simpler than is the
condition in Paragonimus ringeri and Paragonimus kellicolti. In
the former parasite the loops are open, and may be distinguished
readily, but in the latter two forms they are close, and the entire
organ presents the appearance of a solid mass. The age and
stage of development must be a controlling factor in this condition,
and very likely there is no constant difference here between the
species.
It is well known that among the Trematoda one finds at times
an exact reversal in the usual position of the organs, chiefly of the
reproductive system, so that the specimen is a mirror image of the
usual relation. Such a condition has been designated amphitypy,
and occurs in some species so frequently that it is impossible to
say which is the normal and which the reversed situs genitalium.
In Paragonimus such a reversal has been observed by several inves¬
tigators. Ward reported it for Paragonimus kellicotti , and we can
confirm the record. In it the uterus lies on the left side, the right
testis is anterior and slightly larger, the ovary is on the right side,
and the bend in the excretory reservoir is to the right. This
condition is not frequent in this species. Kubo found it in seven
cases out of eighteen in Paragonimus ringeri. It is interesting to
note that in the specimen figured by Looss (1914, p. 321) the organs
are represented in what we regard as the reversed location, and
not in the normal position. One figure published in Leuckart
(1889) showed the same reversal. This has led to some confusion,
139
since the location of organs is stated by different writers in
diametrically opposite terms.
As one of us (Ward, 1908, p. 178) has noted previously, the
correct measurement of ova is not a simple matter. All sorts of
direct errors in counting micrometer spaces and in computing actual
values are not only possible but, in actual practice, frequent. They
are not easy to test or detect. Against the danger of computing
averages from a small number of specimens it is hardly necessary
to warn the student, although this error has been committed by
some experienced men. A more insidious error is caused by the
unconscious selection of the larger and more conspicuous specimens;
in this way an investigator may raise the true average considerably
and reduce the range of size through the elimination of the smaller
specimens. Such a tendency can be detected if the full series of
measurements is given, but can only be inferred with some hazard
if the extreme and average measurements are the only data printed.
The contrary difficulty, which is even more serious and more
difficult to detect, will be introduced by that observer whose
emphasis on the need of reporting every item he sees leads to his
measuring and recording the size of absolutely every egg in a
group. In any preparation one finds a considerable percentage of
ova that are clearly abnormal. The shell is distorted by pressure
or osmotic currents, which have made it over-large or over-wide;
or in its process of manufacture by the parasite some interference
with the normal course of events led to the moulding of an aberrant
shell. The distorted form, or the atypical contents, demonstrate
its unfitness for consideration in such measurements. To measure
and record such an egg in the description of a species, unless the
fact be given in connection with a note on the character of the
individual egg, is usually to hamper, and certainly not to aid,
the work of future students.
Here we may quote again on this matter the views of Looss
expressed in an article in this journal (1907, p. 149) which I cited
previously in my discussion of this topic. He says : —
‘ There exist, of course, among the immense number of ova in
an individual worm always some which are either larger or smaller
than the rest, or even evidently misshapen. In my opinion, it is
of no use to record carefully the measurements of these eggs also.
140
For the description and definition of a species it is much more
important to select for measurement those ova which appear to be
normal, and to present the size and shape typical for the species.
It may be added in passing that young worms with few ova in
their uteri usually do not afford normally-shaped and normally-
sized ova. 1
Finally it does make a difference what is the exact source of
the eggs measured. Those which are taken from the body of the
worm near the beginning of the uterus, i.e., which are just formed,
do not agree in size and proportions with those at the end of the
uterus ready to be laid, or those which are collected after deposition
by the worm. I have noted on several occasions a tendency of the
student to measure ova from the first coils of the uterus, because
there they are less crowded, and hence more easily seen and
measured. With the passage of the eggs through the uterus they
increase in size, probably by imbibition of uterine fluid, and the
increase seems to be most marked in case the ovum enters upon its
development during the intra-uterine period, so that when deposited
the egg shell contains a more or less advanced embryo.
Since one of us (Ward, 1908) has emphasized the significance
of differences in the size of the eggs of Paragontmus as reported
by various observers, especial attention was devoted to a study of
these structures. An effort was made to get series of measure¬
ments under standard conditions, and to ascertain how far these
varied from measurements previously given for eggs from similar
sources. A start was made with Paragontmus kellicotti , and since
it was evident at first thought that eggs taken from the body of the
worm, or measured while still enclosed within it, might differ
from those that had been laid in the normal manner, the first set
of measurements was made from material that had been deposited
naturally by the adult parasite.
The mucous exudate obtained from the bronchi and from the
worm cysts in infected hog lungs was brought carefully into
glycerine jelly and mounted within asphaltum rings. These
preparations contained large numbers of ova, and being protected
from any pressure by the cover-glass, the ova were in a natural,
undistorted and undamaged condition.
Series of these eggs were measured, excluding only such
specimens as were distinctly aberrant in form, or bore evidence of
having suffered some mechanical injury. The maximum length
obtained was 00875 mm. and the minimum o*0775mm., with an
average of 0'o83 mm. The widest specimen measured 0*065 mm.
and the narrowest 0 0525 mm., while the average width was
0*0559 mm.
The only other investigators who have measured these eggs
under similar conditions are Stiles and Hassall, who state
(1900, p. 603) that twenty-five eggs taken from cysts in the lungs
of hogs varied in length from 0*096 mm. in maximum to 0*078 mm.
in minimum, with an average of 0*0856mm., and in width from
006 mm. in maximum to 0*048 mm. in minimum, averaging
0*0532 mm. It will be noticed that so far as the length is
concerned the minimum measurement of Stiles and Hassall is
practically identical with our minimum, but the maximum is nearly
ten micra larger. We were unable to find any eggs of that length
in material mounted so as to remain free from pressure, and it is
clear that they recorded very few since their average size is only
slightly larger than our record. The width given in their records
does not vary greatly from that we record. The slight differences
of from 3 to 5 per cent, in the averages of length and width
can not be regarded as of serious significance. These figures
represent, undoubtedly, the approximate dimensions of the egg
of Paragonimus kellicotti.
In order to determine under similar conditions the size of the
ova from the human lung fluke, slides were made in the same way,
using the sputum obtained from an infected Korean. The material
was not fresh as in the case of the hog parasite, but had been sent
in formol from Chemulpo, Korea. A series of measurements from
this material, after it had been treated exactly like the series of
eggs from the sputum of the pig, gave the following values: —
Length in maximum 0*097 mm., minimum 0*08 mm., with an
average of 00872 mm.; breadth in maximum 0*055 mm.; minimum
0*046 mm., with an average of 0*0506 mm. Some months later we
obtained through the kindness of Dr. M. M. Null, of Seattle,
Washington, sputum from the patient, a Korean also, in that city
who had been found to be infected with the lung fluke (see Null,
M.M., 1910). Four series of eggs from this material, after treat-
142
ment in the same manner as before, were measured separately, with
the following results: —
Length in mm.
Width in mm.
Maximum
Minimum
Average
Maximum
Minimum
Average
Scries (<j) ...
0*0884
0*0754
0*0812
0*0546
0*0442
0*0493
Series (i b ).
0*0884
0*0780
0*0822
0*0520
0*0468
0*0499
Series (r) .
0*0858
0*0780
0*0806
0*0546
0*0442
0*0496
Series (d) .
0*0858
0*0780
0*0813
0
N
»/%
O
6
0*0468
0*0483
General Average
0*0812
0*0492
These series of eggs of the human lung fluke from different
localities do not agree perfectly in measurements. This is
especially noticeable in the length, which in the material from
Korea had a much higher maximum, and consequently an average
about 7 % greater. The width is almost identical, as the range
agrees perfectly, and the average of the one differs only 2% from
that of the other. Furthermore, it will be noted from the measure¬
ments cited that the range of variation in width is slight, being
much less than the range of variation in length. It should be noted,
as explained later, that the form of the egg is identical in the
two cases. By reason of the extreme care used in obtaining these
measurements, we believe they represent closely the true dimensions
of the egg of this species and the probable range of size in normal
undistorted eggs.
There are many other observations on the eggs of the human
lung fluke. In 1880 Baelz reported that the bodies he found in
sputum measured 013 by 0 07 mm. This record may be rejected
as unquestionably erroneous since it does not agree with any other
report, and more especially since it was not cited three years later
by the same author, who then gives the size of the ova as 0*08 to
o' 1 by 0*05 mm. He gives no average size, but this range is close
to that we obtained (0 08 to 0 097 by 0 05 mm.) for the Korean
material. One other record is given for eggs taken fresh from
! 43
human sputum. Manson (1882) records the average size of these
ova as 0*085 by 0*051 mm.
Other records concern measurements made from preserved
material or that of which the condition and method of handling
is not stated. The most aberrant of these comes from Yamagiwa
(1890), who measured in sections of the human brain and lungs ova
which he attributed to the lung fluke, and found them in one case
from 0 04 to 0*064 mm. by 0*024 to 0 04 mm., and in the other from
0*049 to °’°6 mm. by 0*029 to 0*036 mm. These measurements are
so extreme that they can not be accepted as belonging to the same
species as that from which the other measurements were taken. On
the other hand, they agree more closely with the eggs of
Schistosoma japonicum , although smaller even for that species than
the size given by others. Now Schistosoma japonicum has been
discovered since Yamagiwa's work was published, and was not
taken into account in his discussion. Without entering here upon
any discussion of the other species, one may say confidently there is
every reason for eliminating his record from the category of cases
assignable to Paragonimus.
Several other observations which undoubtedly concern the lung
fluke remain for consideration. The record of Leuckart (1889) may
have been made on material from the tiger or on some from the
human host. He gives the length as varying from 0*08 to 0*i mrn.,
which agrees almost exactly with our figures, and states the width
at 0 056 mm. which is barely larger than our recorded maximum
width, 0*055 mm - These are distinctly larger than the figures
given for ova of the tiger lung fluke, and hence probably were
taken from specimens of the human lung parasite. Katsurada
(1900) found the ova of the human lung fluke to vary from 0*0875
to 0*1025 mm. in length with an average of 0*0935 mm., and from
0*0525 to 0*0663 mm. in breadth with an average of 0*057 mm.
Mackenzie (1904) records the length as 0*0855 to 0*0997 mm * with
an average of 0*0913 mm., and the width from 0*048 to 0*069 mm.,
averaging 0*0552 mm. Both the extremes and the averages of these
two records depart rather widely from our observations, and we
are unable at this time to suggest any explanation for the
discrepancy.
Kubo did not devote particular attention to the egg, as he
*44
measured only ten specimens and gave but a very brief description.
According to his records, the average length is 0*07 mm. and the
average breadth 0 06 mm. These figures do not agree even
approximately with those of any other investigator, and show a
shell relatively much shorter and broader than anyone else has
found. In fact, this average length is less than the minimum
figures given in any previous paper, while at the same time the
average breadth is greater than any other average, and equal to
the maximum for this dimension given previously.
These various results obtained from the measurements of eggs
demonstrate wide discrepancies in technic or some inaccuracy in
methods. That the eggs of Paragonimus really vary as much as
these figures seem to indicate is hardly a tenable hypothesis.
Looss, whose accuracy is unquestioned, says of these different
records that he has personally found, in undistorted worms, eggs
only from 0 077 to 0*081 mm. long and from 0 046 to 0*05 mm.
broad, and accordingly the variant records undoubtedly belong to
another species or to much distorted eggs in which the cover is
easily forced off by pressure. We also have been unable to find
any such variation in normal eggs as these figures given by different
writers seem to indicate, even when the eggs are taken from adults
in different hosts or from widely separated regions. Hence we are
forced to assume the introduction of errors of some sort.
In general it appears from these data that the eggs of
Paragonimus kelticotti are broader and shorter than those of
Paragonimus ringeri. These differences are not pronounced, and
are certainly not sufficiently clear to allow of their use in the
differentiation of species, as in extreme cases the sizes overlap.
There is, however, a characteristic difference in form which may be
seen on comparison of the non-operculated ends of the shell. In
Paragonimus kellicotti that end tapers off rather sharply so as to
produce the effect of a pointed extremity. In Paragonimus ringeri
the corresponding end of the shell has a wider curve, due to a
greater flare in the sides, which gives the egg a more pronounced
elliptical outline. While this difference is trifling it is so distinct
that once learned the student can readily pick out the species on
comparison of the eggs. It was noticeable, for instance, that
the two sets of ova from the human lung, discussed on pp. 141-2,
differed somewhat in length from each other, but agreed perfectly
in the outline of the non-operculate end of the shell, so that one
of us measuring them at once recorded this correspondence in his
notes before the data had been compared with other records or any
attempt made to interpret them.
Finally, it is noteworthy that the various records of size
indicate much greater differences in length than in width, and there
is a possibility that the eggs do vary widely in length while
retaining a more uniform standard width. One could readily
frame a hypothesis of egg formation which would agree with the
known structure of the organs and the possible uniformity in width
together with variation in length; but at present these supposed
facts are too uncertain to be used in such a manner.
The eggs of the Philippine forms were very carefully measured
by Musgrave (1907, p. 35), using a Zeiss photomicrographic
apparatus. In length they vary from 0*062 to 0'098 mm., with an
average of 0*074 mm.; and in breadth from 0*047 to 0*063 mm.
with an average of 0*057 mm. Compared with our own measure¬
ments it will be noted that the maximum length is in exact
agreement, but the minimum and the average length are far too
small. It may be, as Looss has suggested, that Musgrave has found
and measured some eggs of Schistosoma due to an intercurrent
infection. The elimination of these lower values will bring the
' minimum and the average into agreement with our figures. But
the width of the eggs as reported by Musgrave is open to the
contrary objection. The minimum value agrees, but the maximum
is far beyond what we found. This, of course, brings the average
up above our record. It is only just to call attention to the fact
that the figures of Musgrave for the width of the eggs agree fairly
with those of the same dimension given by Katsurada and
Mackenzie, although his values for the length of the eggs are very
far below theirs. The evidence is inadequate for a final decision
regarding the Philippine form.
The eggs of the form found in the tiger measure 0*08 by
0*045 mm., according to Kerbert; and Looss, who apparently
re-measured the eggs of the same specimens sent to Leuckart, gives
the dimensions as 0*077 to 0*o8i mm. long by 0*043 to 0*05 mm.
broad. - These figures are practically identical, and as far as they
146
go indicate a slightly smaller egg than is found in the other two
species. We did not have material for a re-examination and
comparative study of these structures. The difference indicated,
even if fully corroborated by a re-study of the material, is too
slight to serve as a diagnostic basis for the species, since as in the
case of the other species maximum values for Paragonimus
westermanii exceed minimum values for Paragonimus ringeri. The
same is true so far as length is concerned for Paragonimus
kellicotti, but the egg of the latter species is always distinctly wider
than the maximum value obtained for Paragonimus westermanii.
The comment of Kerbert, not shown in his figures, that the eggs of
Paragonimus westermanii are slightly flattened at the [both ?]
poles, seems to furnish a slight difference in the form of the egg
between this and the other species.
GENERAL DISCUSSION
The Genus Paragonimus
Looss was the first to give an extended description of the genus,
and his diagnosis slightly modified is reproduced by Stiles and
Hassall (1900, p. 563). The description was so successfully
written that even an extensive study of these three species has
disclosed only minor corrections. Despite their distinctly insigni¬
ficant character it seems worth while to call attention to these
characters. The pharynx, while well developed, manifests a
tendency for a spherical form rather than an elongate, and even in
the extreme case does not depart much from the spherical. In
our opinion the oesophagus should not be designated as very short.
It always appears in total preparations to be shorter than it
actually is, since it rises obliquely towards the dorsal surface and
thus is foreshortened in any longitudinal aspect, while in many
specimens this inclination is so emphasized by contraction of the
body that the branching of the caeca rests directly upon the
pharynx and thus appears to originate from the latter without the
intervention of any median unpaired region which is the oesophagus.
The failure to see the oesophagus in total preparations is often
responsible for the belief that such a region is non-existent when
in lact it is well developed. In such cases it shows up
conspicuously when the living worm stretches the anterior end, and
becomes apparently obliterated when the worm assumes an average
position or is contracted even moderately. In these instances the
true form and relations of the oesophagus may be made out in
lateral view, which one rarely gets of a trematode either naturally
or in preparations, and may also be determined from a study of
serial sections. From these it appears that the oesophagus suggests
often the form of a letter S placed vertically, since it starts
posteriad from a ventrally located pharynx, curves well to the
front, and then turns again towards the posterior end to join the
caeca. The beginning of these branches lies longitudinally so close
to the pharynx that one does not see the vertical separation and
the contracted or twisted oesophagus that joins them.
Looss states that one testis lies obliquely behind the other, and
this is hardly correct in any sense. As we have shown, the central
mass of the one organ is located directly opposite that of the other,
and the only difference is found in the lesser development of the
anterior lobes in one, usually that on the right side. This is
undoubtedly due to the fact that the total mass of the uterus which
lies just anterior to that testis is greater than that of the ovary
which is anterior to the other. When ovary and uterus are
reversed, as has been reported to occur in exceptional cases, then
the testes show the reverse of the usual development in that the
anterior lobes of the left testis are less prominent than those of the
right testis, and the organ appears to be located slightly posterior
to that on the other side. The testes are in fact opposite each other
and one appears slightly anterior to the other only because of the
relative development of the anterior lobes.
Both Looss, and Stiles and Hassall after him, emphasize the
absence of a receptaculum seminis. As has been already shown in
the discussion, this organ is present even though it be only poorly
developed, and the proper form of statement would emphasize this
fact.
As noted above, all of these points are distinctly secondary,
and the genus description remains substantially as outlined by
Looss. On the other hand, one can hardly agree with him in
regarding the genus as nearly related to the Fasciolinae, especially
to Fasciolopsis, as he maintains it to be. Since we have just
148
learned through personal correspondence that Professor Odhner
has in print a discussion of this matter, it seems best to omit here
any elaboration of this topic.
Stiles and Hassall (1900, p. 604) include in the genus two other
species as follows: (A) Paragonimus rudis (Diesing, 1850) from the
lung of a Brazilian otter; (B) Paragonimus comfactus (Cobbold,
1859) from the lung of the Indian ichneumon. Neither of these has
been reported since it was originally found, and of neither have
we been able to secure material for comparison.
Cuticular Spines
It is important to point out more precisely the significance of our
discovery concerning the cuticular spines. They constitute in our
opinion a most convenient and accurate criterion for the distinction
of species, and one which may well be applied to other genera
among Trematoda to the advantage of the taxonomist on the one
hand, and of the practitioner and pathologist on the other. To
the latter, desirous of making a rapid and accurate diagnosis of a
form which falls into his hands, such a definite feature will be of
marked value. This is especially true since no complicated and
time-consuming technic and no array of apparatus are demanded
for the determination. It is sometimes possible to tear off with fine
forceps a piece of the outer skin with some of the spines in situ;
it is always possible to slice off freehand with a razor a thin layer
of surface tissue containing them. Such a fragment, mounted
roughly, gives a good view of the spines, and thus affords means
for diagnosing the species once that the precise character of the
spines has been determined for that species. This is pre-eminently
a method for diagnosis from fresh material, since, as is well
known, in many cases the spines are caducous in life and are easily
lost also if the specimen lies some time in a preserving fluid. In
fact, spines disappear or are overlooked so easily that many
investigators have paid little attention to the statements regarding
their presence or absence given by previous writers, knowing that
the previously described specimens or their owm might easily have
suffered the loss of these structures and yet be in good condition
for the determination of other structural features.
*49
We should not neglect to say that in our experience these
characters cannot be determined in specimens that have been
reduced to series of sections. Several such series of Paragonimus
have been loaned us by colleagues for the purpose of testing the
character of these structures, and in no such case have we been able
to determine anything concerning the precise form or the arrange¬
ment of the spines, even after long and painstaking effort.
Undoubtedly such a series might contain a bit of cuticula, removed
tangentially from a region where the spines were preserved well,
that would give the desired evidence, but after our experience we
are inclined to regard such an occurrence as exceptional, and to
consider serial sections as unfitted to furnish the data desired
regarding the cuticular spines.
We have not been able to find record of a single species in
which the precise form and distribution of these cuticular spines
have been worked out. Yet they evidently possess advantages
afforded by few structural features among the trematodes for the
precise comparison of closely related forms. The great difficulty
in comparing flukes results from the soft and variable form of most
organs and the absence of hard parts. Descriptions are couched in
terms that vary from individual to individual, and measurements
in a given species range beyond the extremes of others, both among
those truly similar and related and also among those that actually
are more distant. The size of the suckers, which has been selected
as one of the usual measurements in specific descriptions, varies
considerably with the contraction of the specimen, and is moreover
not always easy to determine, while it undoubtedly increases with
the age of the specimen, although to what degree has not been fixed.
Similar difficulties in the case of other organs contribute to make
the description of the average trematode a recital of general,
generic and even family characters rather than of specific features.
Various investigators have sought to utilise the ova as characters
for specific differentiation. The difficulties involved in placing
dependence upon this factor are well exemplified by the discussion
of the lung flukes in the preceding pages. Some important
deductions have been made correctly on this basis, but the procedure
is dangerous, as is any such argument based on a single factor.
Furthermore, Looss has shown clearly that there are two species
of Clonorchis in man, and yet the eggs are hardly distinguishable
in size or form.
The demonstration of another anatomical feature composed of
material that cannot be altered by pressure, contraction, or other
mechanical influence, and relatively constant in form among
different individuals, furnishes a diagnostic element of distinct
importance. When our attention was first drawn to the cuticular
spines, and they seemed to furnish such an element, we felt the
matter deserved the most careful study before any announcement
was made. By the work reported in this paper one may fairly
claim that a demonstration has been given for the essential
similarity, even though not for the absolute identity, of these spines
in different regions of the body of a single worm, and also on
different individuals in the same host and in different hosts. We
have been able to recognize a graduation in size and frequence
characteristic of different parts of the body of the worm, and also
a variation in size in different worms which may be due to the
difference in host that harboured the specimens studied, but is more
likely attributable to differences in age and growth of these
structures in different individuals. In spite of the extreme
differences in size and frequence, there is no approachment between
the species, but rather added emphasis upon their real distinctness.
In our opinion, these spines will furnish convenient and precise
specific distinctions between still other species of Trematodes, and
further studies to test this view are now in progress. It may be
pointed out that this would be a most natural condition. Cuticular
outgrowths, such as hairs, spines, scales, and other processes of
varied form, have long been utilized by systematists in other groups
as a means of identifying species, and have proved to be convenient
and accurate characters for the description and differentiation of
species. In animal groups which have been the object of long
and intensive study, and in which taxonomy may justly be said to
be more securely established than in groups of more recent study
and hence less perfectly known, the use of such characters as specific
criteria is generally approved. Even more than that, it may be
said the species are distinguished on the basis of such differences
when otherwise the structure, so far as worked out, is only known
to be identical. Under these circumstances, we do not think we are
venturing on dangerous or untried ground in maintaining the
specific integrity of these three different forms of lung fluke, when
the contention is supported by differences as clear and unmistakable
as those we have demonstrated between the spines. The larger,
more striking morphological differences, which some years back
were regarded as specific in value, are now generally accepted as
of generic or family rank. The proper evaluation of those items
which are confessedly insignificant will yield a firm basis for the
proper conception of species among the lower forms. Nowhere is
there more need of such careful analysis of minor features than
among parasitic worms, where great confusion reigris by virtue of
the rapid and inexact treatment that has been accorded these forms
in the past. Many investigators of the present day have broken
away from that unfortunate tendency, and are analyzing the
structure of such forms with great care. We hope that the factor
to which we have called attention so prominently in this paper may
prove to be of wider usefulness than in this case merely. Even
should that not prove to be true, we are still confident that species
of Paragonimus can be readily and accurately determined by
means of the cuticular spines. Such a precise determination is of
evident general interest, since only by it can be determined the
range of territory infected by a definite form, the number of
different species that threaten man in a given place, and ultimately
the measures that must be taken to eliminate these parasites from
the list of the enemies of man.
152
REFERENCES
Fehleisen, F., and Cooper, C. M. (1910). Paragonimiasis or Parasitic Hemoptysis. Jour.
Am. Med. Assn., LIV, pp. 697-699.
Garrison, P. E., and Lxynes, R. (1909). The development of the Miraadium of Paragonimus
under various Physical Conditions. Philippine Jour. Sci., B, IV, pp. 177-83.
Hanson, H. (1911). Distoma pulmonale in Wisconsin. Johns Hopkins Hosp. Bull., XXII,
pp. 112-4.
Katsurada, F. (1900). Beitrag zur Kenntniss des Distomum toestermanii. Beitr. z. path.
Anat. u. allg. Path., XXVIII, pp. 506-522.
Krrbirt, C. (1881). Beitrag zur Kenntniss der Trematoden. Arch, fur mikr. Anat., XIX,
pp . 5*9-578-
Kubo, N. (1912). Morphologic des Distomum pulmonale. Centralbl. Bakt. u. Par., Abt. 1,
Orig., LXV, pp. 115-138; 13 figg.
Leuckart, Rud. (1889). Die Parasiten des Menschen und die von ihnen herriihrenden Krank-
heiten, 2 Aufl., 1 Abt., 4 Lief., pp. 404-440.
Looss, A. (1899). Weitere Beit rage zur Kenntniss der Trematoden-Fauna Aegyptens.
Zool. Jahrb., Syst., 12: 521-784; 9 pi.
-(1905). Von Wfirmcm und Arthropoden hervorgerufene Erkrankungen. Mense’s
Handb. der Tropenkrankh., I, pp. 77-209; 2 pis.
- (1907). Some Parasites in the Museum of the School of Tropical Medicine, Liverpool.
Annals Trop. Med. and Par., I, pp. 123-152.
- (1914). Wiirmer und die von ihnen hervorgerufenen Erkrankungen. Mense’s
Handb. d. Tropenkrankheiten, II Aufl., II, pp. 311-516.
Mackenzie, A. D. (1904). A case of parasitic hemoptysis or infection with the Distoma
toestermanii. Jour. Am. Med. Assn., XLII, pp. 1133-1135.
Manson, Patrick (1880). Further observations on Micro-Filariae, with Descriptions of
New Species. Jour. Queckett Micr. Club, VI, pp. 138-139.
- (1882). Distoma ringeri and parasitical haemoptysis. China Imperial Maritime
Customs. Medical Reports, XXII, pp. 55-62 ; figs. 1-25.
Musgrave, W. E. (1907). Paragonimiasis in the Philippine Islands. Philippine Jour. Sci.,
B, II, pp. 15-65 ; 11 pl».
Nickerson, W. S. (1911)* Paragonimus in a Cat in Minneapolis. Science, n.s., pp. 33-271.
Null, M. M. (1910). Tochil, or Endemic Hemoptysis. Northwest Medicine, II, pp. 364-366.
Stiles, C. W., and Hamall, A. (1900). The Lung Fluke (. Paragonimus toestermanii) in Swine
and its Relation to Parasitic Haemoptysis in Man. Ann. Rept. Bur. An. Ind., XVI,
pp. 560-611 ; 2 pis.
Ward, Henry B. (1894). Ueber das Vorkommen von Distoma toestermanii in den Vereinigten
Staaten. Centr. f. Bakt. u. Par., XV, pp. 362-364.
-(1903). Data for the Determination of Human Entozoa. Trans. Amer. Mic. Soc.,
XXIV, pp. 103-138 ; 4 pis.
-- (1908). Data for the Determination of Human Entozoa II. Trans. Amer. Mic. Soc.,
XXVIII, pp. 177-202 ; 1 pi.
Yamagiwa, K. (1890). Beitrag zur Aetiologie der Jackson’tchen Epilepsie. Arch. f. path.
Anat. u. Phys., CXIX, pp. 447-460.
* 5 +
EXPLANATION OF PLATES VII-XI
All figures are made from camera drawings of microscopical
preparations, unless otherwise stated.
Plate VII
Fig. i. Paragotiimus kellicotti \ total preparation seen from the
ventral surface. The vitellaria are represented on the
left side of the worm, and omitted from the other side
in order to show ovary, testis, vitelline ducts, and
intestine normally obscured by them. The specimen had
been stained, flattened under pressure, and mounted,
x 7 5 diameters, a Egg from same specimen, x 300
diameters.
Fig. 2. Eggs of Paragotiimus kdlicotti showing ordinary varia¬
tions in form. These specimens were taken from mucus
of lung, x 475.
Fig. 3. Eggs of Paragotiimus kellicotti taken from uterus of
parasite, x 475 diameters.
Fig. 4. Reconstruction of a series of sagittal sections of
Paragotiimus kellicotti showing the left half of the body
in the posterior region as seen from the median sagittal
plane. One can distinguish readily the marginal
vitellaria, the dorsal shell gland with a bit of the main
yolk duct or yolk reservoir, the uterus massed around
the acetabulum, the irregularly-lobed testis with its
vas efferens joining the vas deferens, and finally the
extreme posterior tip of the intestinal caecum of that
side.
156
Plate VIII
Fig s . 5-11. Spines of Paragonimus kellicotti to demonstrate
the substantial identity of these structures from different regions of
the same specimen and pn parasites of the same species obtained
from different hosts.
Fig. 5. From dorsal surface behind oral sucker.
Fig. 6. From centre of dorsum.
Fig. 7. From dorsal surface near posterior extremity.
Fig. 8. From ventral surface between oral and ventral suckers.
Fig. 9. From region near oral sucker.
Fig. 10. From region close to ventral sucker.
Fig. 11. From specimen taken from lung of cat.
Fig. 12. Spines from cuticula of Paragonimus ringeri from lung
of man in Japan.
Fig. 13. Spines from cuticula of type specimen of Paragonimus
westermanii.
Annals Trop. Med . & Parasitol.> Vol. IX
PLATE VIII
C. Tinling & Co., Ltd., Imp .
158
Plate IX
Reconstruction of a specimen of Paragonimus kellicotti from
the pig. The alimentary and reproductive systems alone are
represented. The vitellaria are omitted and also the uterus, except
in fig. 15, where it is outlined in part. The specimen was carefully
preserved, and showed no evidence of distortion. It had not been
flattened under pressure, as have most specimens drawn for illustra¬
tions in the texts. Consequently it shows more accurately the true
form and relations of organs.
Fig. 14. In lateral aspect, showing the right side of the body
only.
Fig. 15. In lateral aspect, showing the left side of the body only.
Fig. 16. In dorsal aspect, showing both sides of the body.
Annals 7rop. Med. & ParasitolVol. IX
PLATE IX
C. Tinting & 1 Co., Ltd., Imp .
i6o
Plate X
Fig. 17. Cross section of intestinal wall, showing character of
epithelial cells and subjacent tissue.
Fig. 18. Cross section of body wall, showing cuticular spines
in situ . The anterior end of the body lies to the right
in the figure.
Fig. 19. Section through ovary and oviduct, including also some
cells of shell gland and the wall of the seminal vesicle.
Fig. 20. Sagittal section supplemented from adjacent sections to
show shell gland complex, including vitelline reservoir
and common yolk duct, oviduct, seminal vesicle, Laurer's
canal, ootype, shell gland, and beginning of uterus.
Plate XI
Fig. 21. Cross section of vas deferens near acetabulum. In
this region the tube is filled with spermatozoa and
surrounded by a loose mass of prostate gland cells.
Fig. 22. Frontal section passing obliquely nearly through the
length of the metraterm, which here contains a single
egg. The layers of the wall and adjoining gland cells
are clearly shown.
Fig. 23. Cross section of oesophagus, showing layers in wall and
associated salivary (?) gland cells.
Annals Trop. Med. ParasitolVol. IX
PLATE XI
C. Ttitling dr 1 Co., Ltd., Imp.
163
PRELIMINARY NOTES ON THE
MOSQUITOS OF KABINDA (LOMAMI),
BELGIAN CONGO
BY
Dr. J. Schwetz
(Received for publication July , 1914)
Kabinda is a healthy station situated at an altitude of about
2,800 feet. Malaria is known to exist, but is somewhat rare and of
little importance. The habitations of the whites are grouped on
the summits of several hills which are separated by ravines of
varying depths. There are no swamps in the immediate
neighbourhood of Kabinda, and mosquitos, in general, are
relatively rare. They are, however, very sensitive to seasonal
variations, and are more numerous at the beginning and end of the
rainy season, when the rains are not excessive.
At the time of my first visit to Kabinda I caught on myself, in
the doctor's quarters, a specimen of Stegomyia fasciata, the presence
of which species had already attracted my attention. Afterwards,
in addition to several 4 ordinary' Culicine mosquitos, I captured a
yellowish species (7 aeniorhynchus) and an Anopheline ( Myzontyia ).
Thus from the beginning I observed that, in spite of the relative
rarity of these flies at Kabinda, representatives of different groups
were present, and that, therefore, an investigation would probably
be of value.
Returning later in order to pass some months at Kabinda,
I settled down to the task of breeding these insects. There was no
difficulty in obtaining the larvae, since it was only necessary for me,
or a trained helper, to make a tour of the European or certain native
compounds. Here would always be found the ‘ reservoirs containing
stagnant water/ so strictly proscribed by malarial prophylaxy,
namely, all kinds of receptacles containing rainwater or water
intended for pigeons, poultry, ducks, etc. These receptacles, which
are scarcely ever emptied, sometimes swarm with mosquito and
Chironomid larvae.
164
Within a few months 1 bred a great number of mosquitos,
several thousands, but before enumerating the tribes and important
genera obtained (I am not sufficiently competent to determine with
certainty the ‘ smaller ’• genera) I wish to make the following state¬
ments : —
(1) I have not been able to breed certain species which, however,
occur in the houses of the station. On the other hand, I have
obtained in this way a mosquito which I have never seen in, or even
near, the houses. Thus, the species of Taeniorhynchus are quite
common in the houses, and I have seen them every evening in my
own; nevertheless, I have not bred a single specimen. The same
may be said for the members of the genus Mansonioides , which,
though very rare, are also to be met with in the houses of Kabinda,
and yet have never been bred. This probably means that the
larvae of Taeniorhynchus and of Mansonioides require a special
type of stagnant water, probably swamp water, and are not able to
live in the more or less filthy water of the receptacles occurring near
human habitations.* A few analogous facts which I have observed
in other localities tend to strengthen this hypothesis. In the
proximity of the station Mutombo- Mukulu there are numerous
marshes, and mosquitos are very prevalent.
During my first visit to this station, the mosquitos captured in
my temporary habitation belonged almost exclusively to the genus
Mansonioides. At the time of my second visit (the station had in
the meantime been removed to a plateau a short distance further
from the marsh), the mosquitos taken in the house were nearly all
the yellowish Taeniorhynchus. During this second sojourn at
Mutombo-Mukulu, however, I bred a certain number of mosquitos,
and yet did not obtain a single specimen of Mansonioides or
Taeniorhynchus. The same facts were observed at Samba
(Kasonga Niembo). On the other hand, I have succeeded in
# The immature stages of Mansonioides have been found by Dr. Ingram, in the Gold
Coast, in borrow pits overgrown with a water plant (Pistia stratiotes). The larvae remain
constantly below, attached, by means of their peculiarly modified siphon tubes, to the roots
of the plant; the pupae, although not remaining continuously below, also attach themselves
to the plant, by grasping the roots with their extremely elongate air-trumpets, and apparently
pass considerable periods in this manner. The larvae of no African species of T aeniorbynebus
have yet been discovered, but the life-histories of three or four American species have been
worked out and are very similar to tho^e of Mansonioides.
H. F. Carter
raising a few examples of Megarhinus>* having found a few larvae
in a receptacle, although these mosquitos are not seen in the
proximity of habitations. This enormous mosquito, with the bent
proboscis, does not occur in the houses, because it does not bite.
But the fact that I have found larvae (four) of Megarhinus* in a
small receptacle containing dirty water and that these larvae
produced adults, shows that this mosquito has no need of special
water or of any particular water-plant.
(2) Generally speaking, the larvae of different species are not
found in the same receptacle in any great variety. Those of a
genus, or even of a species, not only predominate, but often are the
only ones present at a given moment, and therefore the contents of
a receptacle provide, as a rule, only a single species at a time. The
same receptacle has furnished on one day specimens almost entirely
of Culex pipiens,\ and eight days afterwards almost exclusively of
Stegomyia fasciata. This is probably due merely to the chance of
oviposition.
(3) The Anophelines are relatively rare at Kabinda, both in the
adult stage in and near houses, and in the larval stage, in artificial
receptacles containing stagnant water. Among the thousands of
mosquitos which have been bred, only a few dozen were Anophelines;
these were obtained sometimes as rare examples among Culicines,
and on one occasion as a pure ‘ culture 1 (associated Chironomid
larvae having been removed).
Among the Culicines, two important genera, Culex and
Stegomyia , constituted 90 per cent, of those found.
DETERMINATION AND CLASSIFICATION OF MOSQUITOS
CAPTURED AND BRED AT KABINDA
Not being a specialist, I have had and still have great difficulty
in determining the species bred, all the more so since I have been
unable to consult the special works on the subject. The exact
identification is rendered still more difficult owing to the recent
changes, wrought by Mr. Edwards, in the classification and
terminology proposed by Prof. Theobald. As I wished above all
to make this work of value, I resolved to avoid doubtful examples,
# The tribe or sub-family Megar bin ini is intended, not the genus Megarhinus . H. F. C.
t Probably C. iuttoni, Theob. H. F. C.
i66
and have, therefore, confined my determination to specimens which
I recognised. I have sent the others for identification to Mr. F. W.
Edwards (British Museum) and to Prof. R. Newstead and
Mr. H. F. Carter (Liverpool School of Tropical Medicine), and
have asked them at the same time to revise my determinations. I
shall, therefore, be able at a further date to complete and correct
this small preliminary study.
The Culicinae (true mosquitos) are divided into four tribes—
Megarhinini , Sabethini , Culicini , and Anophelini —and representa¬
tives of each have been found at Kabinda.
(1) Megarhinini. In a receptacle containing water for the use
of pigeons, I discovered, on one occasion, among some Culicine
larvae, four enormous larvae which, from a distance, resembled
small fish. Although I had never seen any so large, I immediately
surmised, from their size, that they were probably the larvae of a
Megarhine. Such an opportunity being so rare, I decided to
preserve a larva and pupa; on the appearance of one of the latter
a larva was killed, and when a second pupa developed one of these
was also killed. The two remaining larvae hatched into two superb
Megharines, a male and a female, of the same species, viz. :
Toxorhynchites brevipalpis , Theob. The pupal stages of these
two mosquitos were of considerable duration, and occupied five days
in the case of the female, eight days in the male, whereas this period
in other mosquitos is usually only one or two days.
(2) Sabethini. These mosquitos were found several times in
very small numbers among Culicines; twenty in all have been bred
out, and two specimens were taken in the doctor's house. The
characteristics of the larvae were not observed; all belonged to the
same species— Eretmapodites chrysogaster , Graham.
(3) Anophelini. As previously mentioned, the Anophelines
are uncommon in this district; the few dozen examples raised from
larvae, and the few rare specimens captured, belonged to the genus
Myzomyia , Blanch. The species obtained at Kabinda and at all
the other localities examined in this region—Mutombo-Mukulu,
Samba, Labefu, Pania, etc.—proved to be M. costalis , Loew.
(4) Culicini. This is the commonest tribe of mosquitos but the
richest in genera and species, containing species of very variable
size and colour. In consequence, it is the most difficult group to
study, and the most intricate as regards classification. I have so
far observed representatives of six different genera of the tribe, but
have only been able to determine four with certainty.
(a) Genus Culex , Linn.; C. tigripes , Grand.* Five examples of
this species have been bred, the large larvae being associated with
those of other species of Culex . C. pipiens group t—at least 50 per
cent, of the mosquitos which I bred belonged to this group.
(b) Genus Mansonioides , Theob. As previously mentioned,
the larvae of these mosquitos have not been found, but I have
captured a few adults of M. unifomits ,+ Theob., in the houses.
( c ) Genus T aeniorhynchus % Arr. A score of adults were taken
in my house, although none were bred. All the examples are of the
same species, but show slight variations— T. aurites , Theob. §
(< d ) Genus Stegomyia , Theob. After Culex , my breeding
experiments have furnished me chiefly with members of this genus ;
and almost all my specimens have been found, on very careful
examination, to belong to the famous species, 5 . fasciata , 1 T Fab.
5 . fasciata is very common here, and occurs both in and around the
houses, chiefly towards evening , at which time, after sunset, they
seem to be very active. The following curious fact may be suitably
mentioned here. The mosquitos found in my house belonged,
almost without exception, to the two genera Taeniorhynchus and
Stegomyia . Now while the members of the genus Stegomyia
became active and commenced to bite about sunset, it was almost
exclusively those of Taeniorhynchus which annoyed me later,
between seven and eight p.m.; before this they scarcely ever
appeared.
Besides the four more important genera dealt with above (and
which are of interest in tropical medicine) I have bred numerous
specimens of two other genera of Culicines. Since I am in doubt
• Var. fusca, Theob. H. F. C.
t Judging from the species present in the collection forwarded by Dr. Schwetz, it would
seem that this term refers to C. duttoni , Theob., C. univittatus , Theob., and C. invidiosus,
Theob. ; the true C. pipiens , Linn., was not represented. H. F. C.
t Only a small number of specimens referable to this genus has been received from
Dr. Schwetz, some of which may be Af. ajricanus , Theob. Owing to their poor condition,
however, it is impossible to identify them with certainty. H. F. C.
§ The examples forwarded proved to be T. cristatus , Theob., not T. aurites , Theob.
H. F. C.
H Other species of Stegomyia present in Dr. Schwetz* s collection were S. afrieatta.
Theob. ; S. apieoargentea, Theob.; 5. smpsoni, Theob. ; S. potveri . Theob. II. F. C.
i6H
regarding these, I have sent the specimens for determination to
persons more acquainted with the subject,* and shall refer to them
in my next paper. A certain number of Chironomidae (other than
Ceratopogon) and a few Psychodidae (other than Phlebotomus )
have also been bred, the larvae of these flies being found with those
of the mosquitos. I shall mention these insects again later (as well
as their larvae and pupae) when the specimens have been determined.
My collection of mosquitos has been divided into four parts and
sent to the School of Tropical Medicine, Brussels, the Museum of
Tervueren, the British Museum and the Liverpool School of
Tropical Medicine.
• These specimens are referable to the species Culiciomyia nebulosa, Theob., and
Ochlerotatus ( Protomacleaya) alboventralis , Theob. H. F. C.
169
ON THE PECULIAR MORPHOLOGICAL
APPEARANCES OF A MALARIA
PARASITE
BY
J. W. W. STEPHENS, M.D. (Cantab.)
SIR ALFRED JONES PROFESSOR OF TROPICAL MEDICINE, THE UNIVERSITY OF LIVERPOOI
{Received for publication 30 Nov., 1914)
Plate XII
In a paper entitled ‘A new Malaria parasite of man’ (1914),
I described under the name Plasmodium tenue what I took to be a
new species of malaria parasite. The peculiar morphological
characters described were new to me, and I could find no illustra¬
tions or description of similar appearances.
Since writing the above-mentioned paper I have received through
the kindness of Dr. Le Fanu, W.A.M.S., from the Gold Coast, a
blood slide in which he thought he had recognised the forms I had
described as P. tenue.
Dr Le Fanu had put this slide aside for further study, as he had
noted before my description of P. tenue that there was something
peculiar about the morphology of the parasites. The film was made
from the blood of a native child that came for treatment. The
infection is not a severe one: it is easy to find fields without any
parasites. The forms in Dr. Le Fanu’s slide are, however, in some
respects even more peculiar than the P. tenue forms.
Firstly, I would point out that the blood cells present no evidence
whatsoever of stretching or distortion, and the same peculiar forms
of parasites occur in all parts of the film, thick and thin.
Secondly, there are present large forms of quartan parasites
which appear to me to be quite normal. In the slide it is possible
to trace a transition from normal ring forms to those in which
chromatin particles or strands without any protoplasm occur in the
red cells. A better idea of these will be got from the accompanying
plate than from any description however detailed.
Three views seem to me possible as to the nature of these forms.
(1) That they are a new species of parasite.
170
( 2 ) That they are degenerative, i.e., formed in the body under
unknown conditions, and so perhaps analogous to the so-called
quinine forms of parasites.
(3) That they are artificial, i.c., formed outside the body under
unknown conditions.
The fact that among the parasites figured, forms are found in
which chromatin alone without any protoplasm occurs in the red
cell, is I think in favour of, but not decisive for, one of the latter
two views rather than the first. The fact that normal quartan
parasites are present is against these two latter views. It should
be noted also that forms consisting of protoplasm alone, without
chromatin, were not seen. Nor were pigment grains apart from
parasites seen on the red cell.
While this paper was passing through the press, Balfour and
Wenyon (1914) published a paper entitled ‘The so-called
Plasmodium tenue (Stephens).’ In it they express the opinion that
I had ‘ not produced any evidence to prove that he [I] was not
dealing with an amoeboid sub-tertian parasite.’ The fact that no
observer (with perhaps one exception unknown to me when I wrote)
since Laveran’s discovery of the malaria parasite in 1880, i.e., during
a period of thirty-five years, had described such amoeboid forms of
the malignant tertian parasite, appeared to me to be very strong
evidence against this view.
It is common knowledge that the malignant tertian parasite is to
a certain extent amoeboid, but this is a very different condition from
that I described in P. tenue. The authors quote for instance
Ziemann, but a reference to his coloured plate will make it evident
that he was not describing P. tenue forms. With regard to the
possible exception I mentioned, I much regret that I had
quite overlooked the existence of Dr. Balfour’s plate published in
1908 and reproduced in the paper cited above. To Dr. Balfour
certainly belongs the credit of the first description of markedly
amoeboid forms of malaria parasites (other than the simple tertian),
but a careful comparison of my plate illustrating P. tenue with that
of Dr. Balfour will show that although there is a resemblance, it is
not a close one. Now Ziemann’s plate was published in 1906, and
if extraordinary amoeboid activity of the malignant tertian parasite
was already recognised it seems hardly necessary for Dr. Balfour
to have published his plate of ‘curious amoeboid forms’ in 1908.
But to avoid controversial matter, let us now consider the
substance of Balfour and Wenyon’s paper. They figure two
parasites, the first of which (in a single film from West Africa) is
almost certainly identical with P. tenue, and possibly, too, this is
true of the second parasite (from Bagdad). In this latter case, several
films were taken just before death and an hour previously; all
contained the same forms (Dr. Wenyon in a private letter). As to
the nature of these parasites the authors appear to be in doubt, for
in one part of their paper they state that ‘ it may be possible to find
for these variations some mechanical explanation,’ and in another
that they are ‘young parasites’ (of P. falciparum ) ‘which are
particularly amoeboid for some reason not clearly understood.’
Craig (1914), on the contrary, states that he ‘ is satisfied that
Plasmodium tenue is an atypical form of Plasmodium vivax.'
It is perhaps not out of place to mention here that Emin (1914)
has described a new malaria parasite under the name Plasmodium
vivax, var. minuta.
When we have evidence as to what the nature of the forms I have
described in this paper may be, and particularly whether they do
or do not belong to the large quartan forms, we shall then probably
be in a better position to estimate whether my view that P. tenue
is a new species of malaria parasite is right or wrong.
It is of interest to record the result of an examination of further
films from the case of P. tenue. Two films (March, 1914) showed
quartan, and in one of the films a single pigmented (presumably
simple tertian) parasite was found with the pigment in the form of
rods, in an enlarged cell showing Schuffner’s dots. Three films
(June) showed quartan, and one (July), quartan. Young ring forms
were not found in any of the films.
REFERENCES
Balfoct, A., and Wenyon, C. M. (1914). ‘ The So-called Plumodium tenu* (Stephen.).’
Jour, of Trop. Med. and Hyg., VoL XVII, p. 353. V
Cxaig, C. F. (1914). 1 New varieties and .pecie. of Malaria plawnodia.’ Jour, of Para.it.
(Urbana, Illinois), Vol. I, p. 85.
Emin, A. (1914). ‘ Une vari£t6 nouvelle du parasite de Laveran.* Bull. Soc Path exot
t. VII, p. 385.
Stephens, J.W.W. (1914). * A New Malaria Parasite of Man.’ Proc. Roy. Soc., B. Vol
LXXXVII, pp. 375-377 j and Annals of Trop. Med. and Parasit., Vol VIII
pp. 119-122. ’
7 2
EXPLANATION OF PLATE XII
The figures were drawn with an Abbe camera lucida at a
magnification of 2300 (approximately).
Annals Trot>. Med. £* Parasitol., IW. /.V
P/..-J7A A7/
ON SOME PREVIOUSLY UNDESCRIBED
TABANIDAE FROM AFRICA
BY
HENRY F. CARTER, F.E.S.
(LECTURER IN ENTOMOLOGY, LIVERPOOL SCHOOL OF TROI*ICAL MEDICINE)
{Received for publication I February , 1915)
Plate XIII
The flies considered in this paper belong to the genera Tabanus
and Haematopota of the sub-family Tabaninae; with one exception
they were obtained either from the Transvaal or from the West
Coast of Africa. Those from the Transvaal were all taken in the
vicinity of Onderstepoort, near Pretoria, and formed part of a
collection which also included the following species: Tabanus
sericeiventrisy Loew, T. insignis , Loew, T. taeniola y P. de B.,
T. ditceniatuSy Macq., T. atrimanus, Loew, Haematopota scutellaris y
Loew (?), Chrysops stigmaticaliSy Loew, and Diatomineura aethio -
pica y Thun.
The types and co-types of the various species and of the variety
herein described are in the collection of the Liverpool School of
Tropical Medicine.
Genus TABANUS, Linn.
Tabanus triquetrornatuSy n.sp.
Medium-sized species with the dorsal surface of the thorax and
abdomen chocolate-brown in colour; anterior half and posterior
border of thorax, including whole of scutellum, pale greyish; hind
margins of abdominal tergites with narrow pale borders expanding
in the central region of the second, third, fourth and fifth visible
segments into conspicuous greyish triangles and laterally into
somewhat irregular pale areas. Eyes bare. Legs dark, the tibiae
almost entirely white or creamy white. Wings faintly and uniformly
tinged with brown.
1 74
Head (fig. l): Face, jowls and posterior region steel-grey,
clothed with rather long white hairs; sub-callus (denuded) shining
dark-brown. Front very narrow, from eight to nine times as long
as the width at the base; yellowish-brown in colour, somewhat
darker at the vertex, and clothed with short yellow and black
hairs. Frontal callus shining, dark-brown, rectangular, reaching
from eye to eye, and with a linear prolongation in the central
line extending slightly further than the middle of the front.
Fir,, i. (<i) Tabanus triquetrornatus, n.sp., front view of head, female, (x 13, about).
(1 b ) Antenna of Tabanus triquetrornatus , n.sp., female, (x 13, about).
Antennae (fig. ib) brownish-drab, darker, almost black, at the
tips; first joint clothed with long white and short black hairs;
second joint with a few white hairs ventrally and numerous minute
black ones fringing the apical margins; third joint somewhat
elongate with the dorsal process moderately pronounced and
bearing a few short black hairs. Palpi creamy white, clothed with
white hairs which, on the outer side of the terminal segment
towards the apex, are intermingled with short black ones.
Thorax : Anterior portion of dorsum, as far as the transverse
suture, ashy-grey, clothed with yellow and black hairs, the former
apparently* more numerous laterally; remaining portion of dorsum
The dorsal surface of the thorax is partially denuded in this specimen.
i75
(except area immediately in front of scutellum, basal half of post-
alar calli and scutellum , all of which are yellowish-grey) of a
chocolate-brown colour; humeral and post-alar calli bearing long
white hairs. Pleurae steel-grey, slightly darker anteriorly with
whitish hairs.
Abdomen : Dorsal surface chocolate-brown, clothed with short
black hairs; distal margins of segments, triangular areas and lateral
expansions light grey, with yellowish hairs; pale triangles on
second, third and fourth segments large, their apices reaching
almost to the basal margins of the segments on which they are
situated, that on the fifth segment considerably smaller and only
half the height of the segment. Venter , two basal segments entirely
grey, clothed with white hairs; remaining segments chocolate-brown
with pale grey hind margins expanding somewhat laterally, clothed
with white hairs interspersed on the darker areas, with black ones
which become longer and more numerous on the last two segments.
Wings : Faintly brownish; veins and stigma pale reddish-
brown. Squamae chocolate-brown. Halteres , stems and knobs
brown.
Legs: Coxae grey, clothed with white hairs. Femora black,
more or less pruinose ventrally and laterally, especially those of the
hind legs; dothed with black hairs which are intermingled with,
or replaced by, white hairs on the pruinose areas. Tibiae, except
extreme apices, white or yellowish-white clothed with white hairs.
Apices of tibiae and whole of tarsi black, with short black hairs.
Lengthy 12 mm.; length of wing \o' 2 $ mm., width of head
4 mm.; width of front at vertex o’5 mm.
Habitat: Calabar, Southern Nigeria; Dr. T. B. Adam, 1913;
one female.
This species is evidently related to Tabanus argenteus % Surcouf.
It is easily distinguished, however, by its general paler coloration,
paler wings, and by its abdominal ornamentation.
Tabanus fuscipesy Ric., var. oculipilus n. var.
Closely resembling the typical form, but with the eyes more
thickly covered with longer hairs, the femora distinctly paler and
the basal portion of the third joint of the antenna somewhat shorter
and broader. The femora are yellowish, only the basal third of
i 7 6
the front pair and the extreme bases of the middle and hind pairs
being dark. The recurrent veinlet from the upper branch of the
cubital fork is usually considerably longer and more conspicuous
than in the typical form.
Fig. 2. ( a ) Antenna of Tabantis fuscipes var. oculipilus , n.var., female, (x 35, about).
( 3 ) Antenna of Tabanus fuscipes, Ric., female, (x 35, about).
Length , 11*25 to 13*25 mm.; length of wing, 875 to 10*25 mm -
Habitat : Onderstepoort, Transvaal; type female, 2.2.1913,
Mr. G. A. H. Bedford; thirteen females, presented by Mr. F. V.
Theobald, taken in the same district during January, February and
March, 1911.
Both Tabanus fuscipes , Ric., and its near relation T . ditceniatus ,
Macq., belong to the sub-genus Atylotus , Osten-Sacken, all the
members of which possess pubescent eyes. In these two species,
however, the hairs are almost microscopical, whereas in the new
variety the pubescence of the eyes is very marked and clearly
visible under a pocket lens (x 15). Taking into consideration also
the other differences mentioned it would seem that the form in
question is worthy of varietal rank. T. taeniatus f Macq., which
also occurs in South Africa and possesses hairy eyes, might perhaps
be confused with this insect but may be distinguished, inter alia ,
by the presence of two sharply defined whitish, admedian, stripes
on the thorax, by the abdominal stripes being white instead of
yellowish, and by the legs and antennae being darker.
Genus HAEMATOPOTA, Mcig.
Haematopota transvaalensis , n. sp.
(Plate XIII, fig. 3)
Medium-sized blackish species with conspicuous, pale, more or
less circular spots on each abdominal tergite, grey, clearly-marked
wings and dark legs—the front tibiae strongly incrassate.
i ?7
Head (fig. 3) : Face , jowls and posterior region whitish, clothed
with long white hairs; upper part of face with a row of four
velvety-black spots extending horizontally just below the bases of
the antennae; each of the inner spots is rounded and is situated
immediately below one of the antennae, while each outer spot is
more or less triangular with its base touching the eye; face above
the line of spots yellowish-brown with numerous, irregularly
distributed, small black spots. Front rather dark mouse-grey,
except the extreme lateral margins and round the lateral frontal
spots, where it is much paler and almost white; vertex yellowish-
brown, but with a short, narrow, greyish stripe in the middle line
Fig. 3. Haematopota transvaalensis , n.sp., side view of head, female, (x 18, about).
extending to the median frontal spot; front clothed with black and
white hairs, the latter present on the paler areas. Lateral frontal
spots large, rounded, touching the eyes; median spot small and
inconspicuous. Frontal callus shining, olive-brown, of moderate
depth and extending from eye to eye; spot below callus black, or
nearly so, and conspicuous. Antennae (fig. 3) comparatively long
and slender; first joint cylindrical, dark greyish-brown, clothed
with short black hairs; second joint and extreme base of third,
yellowish-brown, the former clothed with minute black hairs; third
joint, except extreme base, very dark brown, terminal annuli almost
i 7 *
black, the last annulus about the same length as the two preceding
together. Palpi grey; terminal segment clothed with short black •
hairs on outer side, replaced towards the base by a few rather long
whitish hairs.
Thorax \ Dorsal surface dark brown with three longitudinal
light grey or whitish stripes; median stripe narrow, more clearly
defined anteriorly, extending from front margin to scutellum
(excepting a short distance in the centre where it is obliterated) and
dilating somewhat posteriorly; lateral, admedian, stripes broader,
terminating just above the inner extremities of the transverse suture,
below each of which is a conspicuous, more or less triangular, white
spot; side of dorsum and humeral calli darker grey, crescentic
marks on hind margin sharply defined, whitish, each with the base
of an ill-defined, moderately large, smoke-grey triangular area
immediately above its outer extremity Scutellum dark brown; the
front margin yellowish-brown in the centre merging into light grey
laterally, sides and broad median stripe, which extends to hind
margin, grey. Dorsum sparsely clothed with glistening white
hairs, humeral calli with long white hairs, post-alar calli with white
and black hairs intermingled, and scutellum with hairs similar to
those of the main portion of dorsum. Pleurae light grey, clothed
chiefly with white hairs.
Abdomen : Dorsum dark brown, the lateral borders of the
segments grey and the hind margins narrowly creamy-white; each
tergite with a pair of comparatively large, yellowish to grey,
admedian spots, and the third to sixth tergites each with a narrow
grey median line; yellowish-grey hind margin of second tergite
produced into a forwardly directed yellowish triangle. Dorsum
clothed with black or brown hairs on the dark areas and with
glistening white hairs on the remaining portions. Venter steel grey,
with hind margins of segments paler, clothed with glistening white
hairs; last segment darker with some erect black hairs.
Wings : Grey with rather fine pale markings as shown in
Plate XIII, fig. 3; rosettes well-defined, apical sinuous mark
single, extending from costa in first submarginal cell (where it is
expanded) to wing margin at a point just below the lower branch
of the third longitudinal vein, distal angles of third and fifth
posterior cells, each with a large pale blotch. Stigma conspicuous,
179
dark brown with the proximal third paler; veins brown. Squamae
pale, tialteresy stalks cream-coloured, knobs dark brown.
Legs : Coxae dark grey (upper half of front pair paler), clothed
with whitish hairs. Femora mostly pruinose, clothed with white
and black hairs, the former preponderating except on the front
femora where the black hairs are more numerous (especially at the
apices) and rather longer. Front tibiae black, distinctly swollen,
with a conspicuous creamy-white ring at the base, middle and hind
tibiae blackish-brown, each with two cream-coloured rings, hind
pair slightly dilated; clothed with black hairs except on the pale
bands where white or creamy-white hairs are present—on the hind
tibiae the pale hairs extend, for some distance, on to the dark space
between the rings. Tarsi black or dark brown, the first tarsal
segments of the middle and hind pairs of legs mostly cream-coloured.
Lengthy io mm.; length of wing, 9 mm.; width of head, 3*5 mm.;
width of front at vertex, ri mm.
Habitat : Onderstepoort, Transvaal; one female presented by
Mr. F. V. Theobald, 24.3.1911, and one other female by
Mr. G. A. H. Bedford, 9.12.1912.
H . transvaalensis does not seem to resemble closely any of the
species described by Loew in his work on South African Diptera,
but is apparently related to Haematopota masculosifaciesy Aust.
From this, however, it may be distinguished by its larger size,
paler scutellum, differences in wing markings ( inter aliay the apical
sinuous mark), and in its abdominal ornamentation.
Haematopota theobaldiy n. sp.
(Plate XIII, fig. 4)
Medium-sized chocolate-brown coloured species with reddish-
brown wings; dorsum of thorax paler with three longitudinal smoke-
grey stripes, dorsum of abdomen darker with a conspicuous ashy-
grey median stripe and traces of spots on, at least, the apical
segments. First antennal segment much swollen and shining black.
Legs dark, the front tibiae incrassate with one pale ring, mid and
hind tibiae with two pale rings.
Head (fig. 4): Face, jowls, and basioccipital region light grey,
clothed with whitish hairs; upper part of face with a transverse row
i8o
of black spots, as in the preceding species, but less distinct, the
inner ones, below the antennae, reduced to mere dots, the outer ones
narrowly separated from the eyes. Front , clothed with glistening
cream-coloured hairs, dark mouse-grey in the centre, light grey at
the extreme lateral margins and round the lateral frontal spots;
Fio. 4. Hatmatopota tbeobaldi , n.*p., side view of head, female. (x 18, about).
vertex with a large dark brown blotch, which becomes somewhat
yellowish-brown at the margins, divided in the central line by a
narrow grey stripe reaching to the median frontal spot. Lateral
frontal spots, more or less circular, not in contact with the eyes;
median frontal spot much reduced and inconspicuous. Frontal
callus shining black, of moderate depth and extending from eye
to eye; upper margin slightly produced in the centre, lower margin
concave for some distance on each side of the middle line; spot below
callus velvety black. Antennae (fig. 4) dark, moderately long; first
joint strongly incrassated and shining black (greyish pollinose at
base and on inner surfaces), clothed with sparse black hairs; second
joint dull reddish-brown, upper angle somewhat produced, clothed
with black hairs; third joint dark brown, slightly paler at extreme
base, with the terminal annuli black—last annulus not quite so long
as the two preceding taken together. Palpi dark grey; first
segment clothed with white hairs, second segment clothed on the
outer side with short black hairs interspersed with rather longer
white ones towards the base.
Thorax : Dorsum chocolate-brown with three longitudinal, rather
dark smoke-grey lines; median stripe narrow, paler at extreme
anterior end, and extending to the scutellum; admedian stripes
shorter, widening and becoming contiguous with median stripe
anteriorly, terminating immediately behind the transverse suture as
a more or less triangular spot; usual crescentic marks on the hind
margin inconspicuous, sides of dorsum, including humeral calli,
grey. Dorsum sparsely clothed with glistening yellow hairs; humeral
calli with long whitish hairs. Scutellum chocolate-brown with hind
margins and central region paler. Pleurae pale mouse-grey clothed
with white hairs.
Abdomen : Dorsally rather darker than thorax with a relatively
broad ashy-grey median stripe, and paired, more or less circular,
similarly coloured, ill-defined, spots on the second to fifth segments;
hind margins of segments, and part of front margin of second
segment, narrowly ashy-grey, and extreme lateral margins of tergites
one to four, light grey, the pale area becoming gradually smaller
towards the fourth segment. Hairs clothing dorsum similar to
those on thorax, but interspersed with black ones, especially on the
darker areas and on the apical segment; lateral pale markings clothed
with white hairs. Venter pale mouse-grey anteriorly and laterally,
gradually merging into chocolate-brown towards the apex, hind
margins with very narrow pale borders; clothed with short yellowish
hairs proximally, with black hairs distally, and with hairs of either
colour on the central segments.
Wings : Reddish-brown with rather coarse pale markings
arranged as shown in Plate XIII, fig. 4. Stigma y elongate, dark
reddish-brown, the basal third paler, yellowish-brown. Squamae
whitish. Halteres with cream-coloured stems and dark brown
knobs, the latter divided in the middle line by a fine creamy stripe.
Legs : Coxae dark grey; those of the fore legs clothed with short
black hairs and some longer white ones towards the base, those of
the middle and hind legs clothed with short whitish hairs. Femora
dark brown, paler and greyish ventrally, clothed both with black
and white hairs—the latter situated chiefly on the pale areas.
Tibiae dark brown; front pair almost black, strongly incrassated
and with a narrow cream-coloured band at the base; middle and
hind pairs with two rather inconspicuous pale bands; tibiae, except
pale bands, clothed with black hairs, the bands with yellowish-white
hairs. Front tarsi black, clothed with black hairs; middle and
hind tarsi dark brown, the first segment of each mostly cream-
coloured, and remaining segments of hind pair narrowly pale (cream-
buff) at extreme base.
Length , 875-10 mm.; length of wing 74-87 mm.; width of
head 2 6-2'g mm.; width of front at vertex *9-105 mm.
Habitat: Onderstepoort, Transvaal; three females presented
by Mr. F. V. Theobald (to whom I have much pleasure in
dedicating this species), 1911, and one female by Mr. G. A. H.
Bedford, 1913.
Mr. E. E. Austen, of the British Museum, has kindly examined
one of the above specimens, and states that ‘in some respects it
appears to resemble Hacmatopota bistrigata , Loew, but is dis¬
tinguishable at once from that species by inter alia the strongly
incrassated front tibiae.*
Haematopota pinguicornis, n. sp.
Small, dark, strikingly marked species; dorsal surface of thorax
dark brown with a very broad, pale, median, longitudinal area on
the anterior portion and a light coloured posterior margin; dorsum
of abdomen dark brown with narrow greyish hind margins to the
segments. Antennae dark brown, the first joint strongly incrassated
and shining, the third joint short and flattened from side to side.
Wings sepia with very conspicuous pale markings. Legs dark with
pale bands on the tibiae, front pair with one pale ring, middle and
hind pairs each with two pale rings—the hind tibiae somewhat
dilated.
Head (fig. 5): Face y jowls and posterior region, grey, clothed
with white hairs interspersed with a few short black ones; face with
a conspicuous black transverse line extending from eye to eye, and
passing immediately below the bases of the antennae, above this line
the face, except the narrow whitish borders to the eyes, is yellowish-
brown in colour becoming darker towards the antennae and margins.
Front dull chocolate-brown with light grey lateral margins which
extend inwards to enclose, or partially enclose, the lateral frontal
spots and diagonally downwards, from the lateral comers of the
vertex, to encircle the median frontal spot; vertex, between the two
diagonally projecting grey stripes, dark brown but with a fine grey
median stripe extending to the median frontal spot. All three
frontal spots distinct, the median spot rather small and circular,
the lateral spots well-developed and more or less quadrate. Front
somewhat denuded, apparently clothed with black and white hairs,
the white hairs being limited to the paler areas. Frontal callus
shining, dark brown or black, of moderate depth and with a distinct
depression in the middle line; upper margin almost straight and
narrowly separated from the eye at each end, lower margin slightly
convex, its lateral extremities in contact with the eyes. Spot below
callus well-defined, dark brown, quadrate.. Antennae (fig. 5) dark,
clothed on the hair-bearing portions with black hairs; first joint
Fig. 5. Haematopota pinguicornis, n.sp., side view of head,
female, (x 1 8 , about).
much swollen, shining, dark brown but paler (yellowish-brown)
towards the base and on the inner surfaces; third joint short, rather
longer than the first and second joints taken together, with the
expanded portion much compressed laterally and somewhat abruptly
constricted basally; annuli fairly distinct, the last annulus about the
same length as the two preceding together. Palpi grey; first
segment darker and somewhat brownish, clothed chiefly with long
white hairs, a few black ones present towards the apex; second
segment rather narrow, clothed on the outer side with numerous
relatively long black hairs, interspersed with a few white hairs.
Thorax : Pale median area on dorsum extending longitudinally
from anterior margin of thorax to slightly beyond level of transverse
suture, its outer margins (corresponding to the usual admedian
stripes) sharply defined, light grey, gradually darkening and
becoming pale brownish in the central region, in which the median
stripe may be traced as a narrow slightly paler line; light grey outer
margins, immediately above inner lateral extremities of transverse
suture, each curving inwards and uniting, near the middle line, with
the inner edge of the light grey spot, situated below the inner lateral
extremity of the transverse suture; crescentic marks on posterior
margin large, light grey; humeral calli grey with white and black
hairs intermingled, the white preponderating; sides each with an
oval grey area, immediately behind humeral callus extending back¬
wards as a moderately broad line to outer extremity of post-alar
callus where, after a short interruption, it curves inwards and
upwards forming a somewhat conspicuous spot. Scutellum dark
brown, paler towards the upper margin. Dorsum clothed with both
black and white hairs, their distribution corresponding more or less
to the darker or lighter markings. Pleurae dark grey, clothed with
white hairs.
Abdomen: Dorsum dark chocolate-brown, clothed with short black
hairs, the hind and lateral margins of the tergites grey or yellowish-
grey, clothed with white hairs. Venter , first segment greyish-brown,
others with colouring and ornamentation similar to that on the
tergites; clothed with short black hairs on the dark portions and
glistening whitish hairs on the narrow grey margins, last ventral
scute with numerous long black hairs.
Wings (Plate XIII, fig. l) : Dark with sharply defined coarse
pale markings; apical sinuous mark strongly bifurcate, both branches
broad, the outer ramus situated close to the wing margin; posterior
cells, except fourth, and axillary cell with conspicuous pale blotches
in their distal marginal angles, fourth posterior cell with a small
indistinct pale mark in its distal angle; other markings as in the
figure. Stigma somewhat elongate, very dark brown, conspicuous,
with a dark rectangular blotch below extending nearly to upper
margin of discal cell. Squamae white. Halteres , stalks cream-
coloured, knobs dark brown.
Legs : Coxae dark brown, clothed with black hairs, basal half
of front pair grey with long white hairs intermingled with the black
ones. Femora dark brown, almost black, clothed with black hairs,
middle and hind femora rather paler ventrally and laterally, the
hind pair slightly thickened. Tibiae, front pair black, each with
a narrow cream-coloured band near the base, middle and hind pairs
dark brown with two pale rings; whole of tibiae, except on the pale
bands, clothed with black hairs, the pale bands with white hairs.
Tarsi, front pair black, middle and hind pairs dark brown, each
with the first segment mostly cream-coloured.
Lengthy &3 mm.; length of wing 6*8 mm.; width of head
2 5 mm.; width of front at vertex *8 mm.
Habitat: Lorha, Gold Coast; Dr. J. F. Corson, 18.4.1914; one
female.
This prettily marked species does not seem to be closely related
to any West African form.
Haematopota angustifronSy n. sp.
Dark medium-sized species with a relatively narrow front;
dorsum of thorax dark sepia-coloured, with two pale admedian
stripes on the anterior half, light grey posterior border and pale
scutellum; dorsal surface of abdomen dark with traces of paired
admedian spots on the apical segments. Wings warm sepia-
coloured with coarse creamy-yellow markings. Legs dark, tibiae
with the usual pale bands, front tibiae slightly dilated, hind tibiae
uniformly thickened.
Head (fig. 6): Face , jowls and basioccipital region light grey
clothed with glistening yellowish hairs; face with two circular black
spots, one immediately below the base of each antenna. Front
narrow, its length about one and five-eighths times the width at the
vertex, dark velvety brown in colour with very narrow yellowish-
brown lateral margins extending to encircle the frontal spots; vertex
with a darker brown, more or less triangular, blotch divided in the
middle line by a fine pale stripe; front clothed with short black
hairs, sparse golden-brown hairs on the paler areas. Three frontal
spots present, the median small but distinct, the laterals large, some¬
what rhomboidal in shape, and either narrowly separated from or
in contact with the eyes. Frontal callus deep, shining, almost
black, not extending from eye to eye, prominent; upper margin
rounded but slightly depressed on each side of the middle line,
lower margin concave above the antennae. Dark spot below callus,
186
quadrate, conspicuous. Antennae (fig. 6 j first joint dark yellow
becoming yellowish-grey towards the apex, long and cylindrical,
not incrassate, clothed with black hairs; second joint similarly
coloured, its upper angle very slightly produced ; third joint
elongate, dark brown to dark yellowish-brown, paler, yellowish-
brown at extreme base; annuli distinct, almost black, the third very
Fig. 6. Hatmatopota angustifrons , n.sp., tide view of head,
female, (x 18, about).
little shorter than the two preceding taken together. Palpi first
segment greyish-yellow, clothed with long glistening cream-coloured
hairs ventrally and laterally, and sparsely with long black hairs
dorsally; second segment, except a large area on the outer side,
greyish-yellow, clothed with long cream-coloured hairs, outer side
mostly dark grey covered with short black hairs.
Thorax: Admedian stripes on dorsum, creamy white, extending
nearly to the inner extremities of the transverse suture, each followed
immediately below suture by a conspicuous, triangular, similarly
coloured, spot; crescentic marks on posterior margin, creamy-white,
sharply defined and relatively large; dorsum clothed with golden-
yellow and minute black hairs intermixed, those of the former colour
more numerous on the sides of the anterior region and longer and
denser on the posterior region; humeral calli mouse-grey clothed
with long whitish hairs. Scutellum creamy-white with dark hind
border, the pale area prolonged in the middle line in some specimens
to the apex; clothed with creamy-white hairs. Pleurae greyish-
brown above, slate-grey below, clothed with pale yellowish hairs.
Abdomen: Slightly darker than the thorax with yellowish hind
margins to the segments and a pair of ill-defined, elongated, dull
yellowish, admedian spots on the fourth, fifth and sixth tergites;
clothed with short black hairs on the main portion, yellowish hairs
on the paler hind margins and rather long, dense, glistening white
or yellowish-white hairs on the extreme lateral apical corners.
Venter greyish-brown darkening towards the apex, with hind
margins yellow but becoming greyish laterally; clothed with
glistening yellowish hairs interspersed with black hairs in the central
region of the penultimate segment, and with erect black hairs on the
last segment.
Wings (Plate XIII, fig. 2) : Pale markings coarse, creamy-yellow
and conspicuous; rosettes large and distinct; apical sinuous mark
relatively broad in its upper and lower portions but greatly
narrowed and sometimes interrupted near the middle of the apical
cell; posterior cells, except the fourth, With very large pale blotches
along the wing margin, that in the first being formed by the lower
extremity of the apical mark; axillary cell with a broad clearly-
defined pale band running from the basal loop round the proximal
margin and joining a somewhat sinuous pale line which extends
upwards to the fifth longitudinal vein, and with a large pale blotch
in its distal angle which extends well into the proximal angle of the
fifth posterior cell. Stigma elongate, conspicuous, brownish-black,
the extreme basal portion yellowish; veins brownish-black. Squamae
brown. Halteres entirely pale cream-coloured.
Legs: Front pair black, middle and hind pairs dark brown.
Coxae of the first pair of legs dark brown or black with the proximal
fourth greyish; clothed with long white hairs basally and with black
hairs on the remaining portion. Femora, front pair clothed with
black hairs which are rather long on the dorsal surface; middle and
hind pairs clothed with black and occasional white hairs, those on
the ventral surface of the hind pair being somewhat longer and more
noticeable than usual. Front tibia with a broad white basal band,
middle and hind tibiae each with two yellowish bands, the distal
bands on the hind pair being generally incomplete, narrower and
188
darker; greater part of tibiae clothed with dark hairs, pale bands
with white or yellowish hairs, interspersed on the hind pair with
black hairs—dorsal surface of hind tibiae with rather long dense
black hairs. Tarsi of middle and hind legs with the first segments
narrowly cream-coloured at the base; clothed with black hairs.
Length , 8‘8-io mm.; length of wing 8*5-9 nun.; width of head
3*25-3*5 mm.; width of front at vertex ’65-75 mm.
Habitat: Belgian Congo; Dr. E. Dutton and Dr. J. L. Todd,
1904; six females.
The exact position of H. angustifrons is somewhat difficult to
define. In a general way it resembles H. grahami , Aust., and
displays affinities to this species in the narrowness of the front, the
shape of the frontal callus and, to a certain extent, in the arrange¬
ment of the pale markings on the body. The most striking character
of H . grahami , however, namely the presence of fringes of long hair
on the femora and hind tibiae, is not present, although the hairs on
these parts in H . angustifrons are perhaps more conspicuous than
usual.
Haematopota exiguicornuta , n. sp.
Small obscurely marked species with extremely short antennae;
dorsal surface of thorax dull brown with three ill-defined, pale,
longitudinal stripes; dorsum of abdomen slightly darker than that
of the thorax, with inconspicuous admedian paired spots on some
of the segments. Legs somewhat pale, the tibiae ringed. Wings
with rather coarse pale markings and inconspicuous stigma.
Head (fig. 7): Face y jowls and basioccipital region light grey,
clothed with white hairs; upper part of face, at the side of each
antenna, dark brown with a narrow black lower border and with
yellowish pubescence. Front sepia-coloured, slightly paler round
the lateral frontal spots and along the margins of the eyes; clothed
with short dark brown hairs. All the frontal spots present but the
median spot much reduced and scarcely visible under the pocket
lens, lateral frontal spots separated from the eye on each side,
moderately conspicuous, and more or less diamond-shaped, each
with its longer axis pointing inwards and downwards. Frontal
callus in the form of a narrow transverse dark reddish-brown band
extending nearly from eye to eye; upper and lower margins gently
curved and almost parallel, the former slightly concave. Spot
189
below callus small dark brown and divided in the middle line.
Antennae (fig. 7) pale, shorter than the head; first joint short,
dark reddish-brown, partly shining and swollen, clothed with black
hairs; second joint paler brown, its upper angle considerably
prdduced; third joint very short, yellowish, considerably flattened
from side to side, the basal portion almost as broad as long;
annuli (at least the first and second) indistinctly separated, the last
not as long as the two preceding taken together and darker brown
H.S.L.
Fig. 7. Haematopota exiguicornuta , n.sp., side view of head,
female, (x 18, about).
at apex. Palpi mouse-grey; first segment clothed with long white
hairs, except dorsally and apically, where black hairs occur; second
segment somewhat dilated basally and gradually tapering to a
bluntly rounded apex, clothed with long glistening white hairs on
the ventral surface, and on the outer side with shorter black hairs.
Thorax : Longitudinal greyish stripes on dorsal surface
inconspicuous, the median stripe extending nearly to the scutellum,
the admedian stripes reaching almost to the transverse suture, each
with an indication of the usual spot below its posterior extremity;
crescentic marks on posterior margin faintly marked, hardly
recognisable; sides slightly paler; dorsum sparsely clothed with
short glistening white and black hairs intermixed. Scutellum dull
brown with greyish hii\d margin, clothed with hairs similar to those
on the main portion of the dorsum. Pleurae grey, clothed with
glistening whitish hairs.
Abdomen : Hind and lateral margins of tergites narrowly light
grey, the pale area being comparatively broad on the sides of the
first and second segments; ill-defined admedian spots present on
the second to seventh tergites inclusive; dorsum clothed mainly
190
with black hairs, pale margins with white hairs. Venter similar
to dorsum, but without admedian spots, with the first segment grey
and with the white hairs more widely and irregularly distributed.
Wings : Pale sepia, the light markings somewhat indistinct and
arranged as shown in Plate XIII, fig. 5. Stigma very faintly
marked, the distal two-thirds pale brown, the proximal third
whitish; veins reddish-brown. Squamae sepia-coloured. Halteres>
stems cream-coloured, knobs dark brown with a relatively broad
creamy stripe along the middle line.
Legs : Pale brown. Coxae grey, clothed on the apex with
black hairs and on the remaining portion with white hairs. Femora
mostly greyish pollinose, clothed with black hairs, interspersed,
especially ventrally and laterally, with white hairs. Tibiae pale
brown, the front pair with a pale basal ring, the middle and hind
pairs each with two pale rings; clothed chiefly with black
hairs, sparse white hairs (which are more numerous on the hind
tibiae) being present on the pale rings. Tarsi, front pair dark
brown, middle pair yellowish-brown, hind pair rather paler than
front pair, each with the base of the first segment yellowish; all
clothed with black hairs.
Lengthy 6 mm.; length of wing, 6*2 mm.; width of head,
2 5 mm.; width of front at vertex, 0*7 mm.
Habitat : Parade ground, Lokoja, N. Nigeria; Dr. W. M.
Manuk, 19.7.1910; one female.
The peculiar and characteristic antennae of H. exiguicornuta
render its identification, and separation from species which resemble
it in general facies, a comparatively simple matter.
Haematopota corsoniy n. sp.
Small dull sepia-coloured species with short antennae and
relatively large dark areas on the face; dorsum of abdomen with
indications of paired spots and with narrow pale hind margins to
the segments. Wings grey, the pale markings somewhat reduced
and inconspicuous; stigma well marked. Legs pale brown, tibiae
unbanded.
Head (fig. 8) : Face light grey with a conspicuous black area
extending between the antenna and the eye on each side, narrowly
separated at its extremities from both, and with the greater part
of the central portion, below the antennae, dark-brown; jowls and
basioccipital region light grey, clothed with long whitish hairs.
Front , sepia with narrow yellowish-grey lateral margins extending
partly round the lateral frontal spots. Median frontal spot absent
or nearly so, lateral spots more or less quadrate and in contact with
the eyes. Frontal callus narrow, shining, reddish-brown, almost
divided by a longitudinal depression in the middle line and with
the lateral extremities of its lower margin touching the eyes; upper
margin nearly straight, curving downwards for a short distance in the
centre, lower margin slightly concave above the base of each antenna.
Spot below callus well-defined, dark brown, triangular, its apex
directed downwards. Antennae (fig. 8) dark yellowish-brown, the
Fig. 8. Haematopota corsoni , n.sp., side view of head,
female, (x 18, about).
tips (annuli) dark brown; first joint slightly incrassate and shining,
clothed with short black hairs; second joint with the upper angle
moderately produced; expanded portion of third joint short—
slightly longer than broad—and somewhat flattened from side to
side, terminal annuli taken together as long, or almost as long, as
the expanded portion, last annulus equal in length to the first and
second combined. Palpi grey, the outer side of the second segment
with short black hairs.
Thorax : Dorsum dull sepia, with three inconspicuous, smoke-
grey, longitudinal stripes extending from the anterior margin to
the region of the transverse suture, the spots below the posterior
extremities of the admedian stripes very faintly marked; crescentic
marks absent; sides, except a small area immediately behind each
humeral callus, and scutellum dark; humeral calli and areas
indicated grey, the former clothed with long white hairs; dorsal
surface sparsely clothed with short whitish or yellowish-white
hairs. Pleurae grey, clothed with white hairs.
192
Abdomen : Dorsum slightly darker than the jthorax, the tergites
with very narrow brownish-grey hind margins, that on the second
expanding into a small greyish triangle in the middle line; lateral
margins of first to fifth segments greyish; paired, admedian spots
on the second to sixth tergites, indistinct, brownish-grey in colour
and approximated to the anterior margins. Venter generally rather
darker than the dorsum, but with the whole of the first and second
segments, except the central portion of the latter, mouse-grey; hind
and lateral margins paler, the hind margins of the proximal scutes
light grey. Both surfaces of abdomen clothed with white and black
hairs, the white hairs limited to the pale margins of the tergites,
but more widely and irregularly distributed on the stemites.
Wings (Plate XIII, fig. 6): Pale markings white, inconspicuous,
distributed as shown in the illustration. Stigma sepia, the extreme
base pale. Squamae white, becoming yellowish towards the
margins. Halteres , stems mostly cream-coloured, but darker at the
base, knobs dark brown with a yellowish-brown central stripe.
Legs : Coxae steel-grey, clothed with glistening white hairs.
Femora, front and hind pairs dull pale-brown, middle pair some¬
what yellowish-brown; clothed with short black and white hairs
intermingled. Tibiae similar in colour to the femora, without the
usual pale rings but with a few short white hairs present on those
positions where the pale bands generally occur, thereby giving the
tibiae faint indications of bands. Tarsi, front pair slightly darker
than the middle and hind pairs, the first segment of each of the
middle and hind pairs faintly yellowish or yellowish-brown at the
base.
Lengthy 5*4 mm.; length of wing, 5 5 mm.; width of head,
2 mm.; width of front at vertex, o*6 mm.
Habitat : Salaga (15 miles North), Gold Coast; Dr. J. F.
Corson (per Mrs. J. F. Corson), 17.10.1914; one female.
This small and obscure species may, on a casual examination,
easily be confused with Haematopota exiguicornutay n. sp. It may
be readily distinguished, however, by the less swollen first antennal
segment and the longer and more distinctly separated terminal
annuli of the third joint, by the more conspicuous stigma and
different arrangement of the pale markings on the wings, and by
the unbanded tibiae, as well as by certain other, less obvious,
differences.
*93
WEST AFRICAN SPECIES OF HABMATOPOTA
Since three of the species of Haematopota described above were
obtained in West Africa, a tabular statement of the more important
distinctive characters of those members of the genus recorded from
the British Possessions on this coast may prove of some value to
medical officers and others interested in the biting flies of this
region. The following table, therefore, refers to the females of those
species at present known to occur in Gambia, Sierra Leone, Gold
Coast or Nigeria, and its preparation has been greatly facilitated by
Mr. E. E. Austen’s excellent descriptions of his numerous species.
1. Front and hind femora and tibiae fringed with long coarse hair ... 2
Front and hind femora and tibiae not fringed with long coarse
hair. 3
2. Hind tibiae greatly swollen with two ill-defined pale rings; frontal
callus widely separated from the eyes; grey thoracic markings
extensive . bullatifrons, Aust.
Hind tibiae scarcely swollen with one broad pale ring; frontal
callus almost touching the eyes; grey thoracic markings not
extensive . grahami , Aust.
3. Legs ornate with pale rings on the tibiae (distinct on, at least, the
front pair). 4
Legs inornate or with obscure rings on the tibiae. 12
4. Hind tibiae with one pale ring . 5
Hind tibiae with two pale rings . 6
5. Blackish species with extensive grey thoracic markings, conspicuously
banded tibiae and pronounced milky-white wing markings
decora. Walk.
Dusky species with dark wings, narrow' grey thoracic markings and
less distinctly banded tibiae . hastata , Aust.
6. Anterior tibiae with two pale rings (the distal ring often faint) ... 7
Anterior tibiae with one pale ring . 8
7. Larger (10-12 mm.) reddish-brown species; first antennal segment
distinctly swollen; abdomen with a rather broad median stripe, and
with trace* of spots on the terminal segments ... vitlata , Loew.
Smaller (9*6 mm.) sepia-coloured species; first antennal segment not
swollen; abdomen with a narrow median stripe and large spots on
all segments. puntens, Aust.
8 . Antennae longer than the head . 9
Antennae very short and inconspicuous, shorter than the head (basal
portion of third joint greatly expanded) ... exiguicornuta , n.sp.
i 9 4
9- First joint of antenna not distinctly swollen . io
First joint of antenna distinctly swollen and shining, small dark species
with conspicuously-marked wings. pinguicomis , n.sp.
10. Brown species; background of wings dark, the light markings
distinct .11
Greyish species; background of wings very pale, the light markings
indistinct . fallidipmnis , Aust.
11. Scutellum brown; first joint of hind tarsi partly yellowish.
torqums , Aust.
Scutellum grey ; first joint of hind tarsi entirely brown.
cordigera , Bigot.
12. Brownish-yellow or greyish species with conspicuous stripes on the
thorax .13
Dark brown species without conspicuous stripes on the thorax (but cf.
tcnuicrus, Aust.) .15
13. Central part of wing hyaline or semi-hyaline, only the tips and hind
border infuscated and with inconspicuous pale markings ... 14
Wings sepia with clearly defined pale markings, extending to the base
pertintns y Aust.
14. Brownish-yellow species ; frontal callus yellowish ... beringcri, Aust.
Greyish species; frontal callus dark brown ... semiclara , Aust.
15. Anterior tibiae not dilated .16
Anterior tibiae distinctly dilated; small dark species with first joint of
antenna swollen . lacessens , Aust.
16. Antennae moderately elongate ; basal portion of third joint slender 17
Antennae very short, scarcely longer than the head; basal portion of
third joint broad . corsoni , n.sp.
17. Thorax conspicuously striped, abdomen indistinctly spotted; first
antennal segment very short and somewhat swollen... tenuicrus, Aust.
Thorax inconspicuously striped, abdomen unspotted; first antennal
segment longer and not swollen. gracilis , Aust.
*95
REFERENCES
Austen, E. E. (1909). Illustrations of African Blood-Sucldng Flics.
- (1908 and 1911). New African Phlebotomic Diptcra in the British Museum
(Natural History), Parts I, II, III, IV and VIII Tabanidae. Ann. Mag. Nat. Hist.,
8th Series, Vol. I, pp. 209 and 401; Vol. II, pp. 94 and 274; Vol. IX, p. 353.
- (1912 and 1914). New African Tabanidae, Parts II, III, IV. Bull. Ent. Res.,
Vol. Ill, pp. 329 and 399; Vol. IV, p. 283.
Loew, H. (i860). Dipt.-Fauna Sudafrika’s, Part I.
Ricardo, Miss G. (1906). Notes on the genus Haematopota of the family Tabanidae in
in the British Museum Collection. Ann. Mag. Nat. Ilist., 7th Series, Vol. XVIII,
P* 94 -
Speiser, P. (1910). Orthorhapha. Zool. Kilimandj.-Meru Exped., 1905-6. Diptcra,
Band II, Abt. 10, p. 31.
Surcoup, Jacques. (1908). Description de Diptires piqueurs Africains, Tabanides du genre
Haematopota. Bull. Mus. Nation. d’Hist. Natur., Vol. XIV, p. 153. Description
d’un Tabanidae (Haematopota) d’Abyssinie. Ibid., p. 224.
- (1909). Note pr 61 iminaire sur la syst&natique du genre Chrysozona. Description
de deux genres nouveaux. Ibid., Vol. XV, p. 453. Description de Tabanides
nouveaux. Ibid., p. 537.
Surcouf, J. M. R., and Ricardo, Miss G. (1909). Etude Monographique des Tabanides
d’Afrique.
Surcouf, J. M. R. (1911-1912). Note sur les Tabanides du Congo Beige des Mue6cs de
Bruxelles et de Tervueren. Rev. Zool. Afric., Vol. I, p. 22. Deuxi£me note sur les
diptires piqueurs du Mus6e du Congo Beige. Ibid., p. 86.
196
EXPLANATION OF PLATE XIII
Fig. r. Wing of Haematopota pinguicornis, n. sp., female (x 7A).
Fig. 2. Wing of Haematopota angustifrons, n. sp., female (x 7).
Fig. 3. Haematopota transvaalertsis, n. sp., female (x 3).
Fig. 4. Haematopota theobaldi, n. sp., female (x 3).
Fig. 5. Wing of Haematopota exiguicormtta, n.sp., female ( x 7-J).
Fig. 6. Wing of Haematopota corsoni, n. sp., female (x 7^).
NOTE.— In figs. 1, 2, 5 and 6 the background of the wings has
been darkened in order to render the light markings more
conspicuous and to overcome halation.
V*
Annals Trop. Med. of Parasitol Fo/. /A'
PLATE XIII
i
2
C. Tinling Co., Ltd. Imp.
'97
EXPERIMENTS WITH SALVARSAN-
COPPER IN TRYPANOSOMIASIS
BY
HARALD SEIDELIN, M.D.
(Received for publication , 23 February , 1915)
With Table
Just before I left for West Africa, in July, 1913, I received from
Geheimrat Ehrlich a supply of a new preparation, K 3 , salvarsan-
copper, which he asked me to use in the treatment of yellow fever
cases. Unfortunately I had no opportunity of complying with this
request, because I saw only yellow fever cases of a mild type, in
which an indifferent treatment was obviously all that was required.
Several severe cases occurred during my stay in Lagos, but they
were either convalescent when they came under my observation, or
they died almost immediately after admission to hospital.
It was also my intention to try the treatment in experimental
yellow fever infection in guinea-pigs, but here again the conditions
were unfavourable. The severity of the experimental infection
varied considerably, and it was never possible to tell whether an
animal would survive or succumb to the disease. Experiments of
this nature would have to be conducted with large parallel series of
infected and non-infected animals, and such experiments we were not
in a position to undertake.
Under these circumstances I resolved to test the value of K 3 in
trypanosome-infections. For this purpose, Dr. J. W. Scott Macfie
kindly supplied me with a strain of a trypanosome ( T . brucei group)
which he had for some time kept in guinea-pigs and rats.
The results of my experiments are given in the accompanying
table, which requires but little explanation. The animals experi¬
mented upon were white rats; the infection was transmitted by means
of intraperitoneal inject tons of a few drops of blood drawn from the
cavernous sinus, according to the technique described by Pettit
(1913). The trypanosome strain proved very virulent, the infected
animals, when not treated, dying from eight to seventeen days after
inoculation.
Geheimrat Ehrlich had advised me to start with doses of 0 05 or
198
o io gramme in experiments upon human patients. The average
weight of the rats was about 60 grammes, and the variations in
weight were small; thus, the proportion in weight of rat to man
would be approximately 1 : 1,000. Accordingly, in the first
experiment I took 0*0001 as a medium dose, using in addition two
higher and two lower doses. As no satisfactory result was obtained
in this experiment I increased the dosage, always proceeding by
multiples of 2. It will be seen from the table that no striking result
was obtained until I reached a dose of eight times the one recom¬
mended, and that the best results, as far as the prolonged absence
of the parasites from the blood was concerned, were obtained by the
injection of 0*0064 gramme instead of 0 0001 gramme. In such a
case the trypanosomes, which had been very numerous before the
injection, were absent from the blood on the following day and
remained absent for fifteen days; the animal died twenty-eight days
after the inoculation. In several other cases I succeeded in
prolonging the life of the animals for a few days more, although the
parasite-free interval was of shorter duration. It appears probable
that repeated injections of smaller quantities would have given better
results, but this method of treatment was not fully investigated. The
number of animals available was not sufficient for work on different
lines, and I therefore confined myself to the investigation, in the
first instance, of what I regarded as the problem of primary
importance, namely, the possibility of establishing a real ‘ therapia
sterilisans magna' In this I failed : a single injection, however
large, did not kill all the parasites in an infected animal, though it
made them disappear, for a considerable length of time, from the
peripheral blood. It would hardly be possible, even in experiments
on animals, to use larger doses- than those I employed, as several
animals died a comparatively short time after the injection without
reappearance of the parasites, and in others a more or less extensive
necrosis of the tissues was produced at the site of injection.
The results may be discussed in a few words.
In series A very little effect was obtained. No appreciable
numerical reduction of the parasites was observed, even after the
injection of 00004 gramme K 3 , but in all cases the parasites
appeared much less lively on the day following the injection than
they had been the day before and as compared with the parasites in
199
the blood of the control animals on the same day. The longest
period of survival obtained in the two animals which had received
the largest doses of K a .
In series B the action of the drug was evident. Two animals
survived the inoculation for 28 and 29 days, respectively, and in one
of them trypanosomes were absent from the blood for six days; this
animal had received an injection of 0 0016 gramme K 3 .
In the two previous series only the doses administered had been
varied, but in series C the drug was injected into animals which
were in different stages of infection. In three animals, as in all
those belonging to the series A and B, trypanosomes were numerous
at the time of injection, but in the other three they had not yet
appeared in the peripheral blood; one animal of the latter group had
been inoculated about 75 minutes before the therapeutical
injection. The stage of infection proved to be of little importance.
The hope had been entertained that a dose, which showed a marked
effect when numerous trypanosomes were present in the blood, might
entirely destroy an infection in its initial stage; but this hope was
disappointed. In rat 232, trypanosomes did not appear in the
peripheral blood until eight days after inoculation; this free interval
may be assumed to be composed of two periods, namely, one
corresponding to the action of K 3 , followed by another corresponding
to the stage of incubation. The dose administered was 0 0016
gramme. The same dose was given in the cases 199, 220, and 229,
in which cases the parasite-free intervals varied from four to six
days. Thus, if we reckon five days for the first period in rat 232,
we get three days for the second period, which is precisely the usual
duration of the incubation stage. In the case of rat 221, trypano¬
somes reappeared in the peripheral blood on the tenth day after the
injection of 0 0032 gramme K 3 ; on the same day 0 0064 gramme was
injected, whereupon the trypanosomes again disappeared from the
blood and remained absent until the animal died, twenty-three days
after inoculation.
In series D the doses were increased. The parasite-free intervals
were prolonged, the maximum of fifteen days after a single
injection being attained in the case of rat 241, which received 0 0064
gramme K 3 ; but the animals did not survive as long as those in
experiment C.
200
In series E repeated injections were given, the dose in each
instance being 0 0064 gramme; in this way a parasite-free period of
twenty days was obtained, but the length of survival was not
increased.
In several of the experiments of the series D and E the animals
died without the trypanosomes having reappeared in the blood.
The experiments in series F were undertaken in order to test the
possibility of producing a K 3 -resistant strain of the trypanosome.
Two rats were inoculated with blood from an animal in which the
parasites had first disappeared after the injection of K 3 , and after¬
wards reappeared. One of the animals was injected twice with K 3 ,
and the effect of the drug was considerably less than in the other
experiments.
The results here reported are interesting from a general point of
view, and leave no possible doubt as to the powerful action of
salvarsan-copper upon this strain of trypanosome, but they are not
conclusive with regard to the therapeutical value of the drug. After
having concluded my experiments, I became acquainted with the
paper by Van den Branden (1913) on the effect of K 3 in human
trypanosomiasis. His results are far superior to those obtained by
me in rats and with a different species of Trypanosoma \ this confirms
the information given me by Professor Ehrlich that K 3 has a
markedly different effect upon different trypanosomes.
The experiments here recorded were carried out whilst I was
working for the Yellow Fever (West Africa) Commission of the
Colonial Office, at the Medical Research Institute, at Yaba, near
Lagos, Nigeria. To the Director of the Institute, Dr. A. Connal,
I am indebted for the facilities afforded me.
REFERENCES
Branden, F. van den (1913). Note prelimimire sur quelques cjsais de traitement dc la
Trypanose Humaine par Salvarsankupfer. Arcb. f. Scb. u. Tropen-Hyg ., Leipzig,
Dec., XVII, 24, pp. 845-849.
Pettit, A. (*913)* Proc&te simple pour pr£lever du sang cher les pstit* rongeurs.
C. R . Soc. Biol., Paris, 10 jan., LXXIV, 1, pp. 11-12.
Result
Date
inoct
[0. 1
tio
1
1 9 *-
1
1611
2: .S
2Z>
1
163 |
22.?
1
164 |
| 21.?
169
I 22.?
165
| 22.)
l66
1^
167
22.1
(68
22.1
*59
, 1 ill
i6e
, Hi
*7
18
*9
Death 13 days after inoculation
i> 7 n
n *4 m
*i 8
» 11 »
u 13 »
99 14 n
11 *7 9*
n *7 »>
» 7 n
n 14 99
Spleen very much enlarged
Peritoneal infection
Spleen very much enlarged
220 |
28.
221
1 28,
229 L
230 Z
23» 3
2 3 2 4
Death 12 days after inoculation j
yy
*3
yy
n
*3
yy
yy
29
yy
yy
28
**
yy
yy
*5
yy
Death 28 days after inoculation
?? -3
M 22
yy 3 2 1? yy
>5 33 v V
91 22 „
[p.T.O.
200
In series E repeated injections were given, the dose in each
instance being 0 0064 gramme; in this way a parasite-free period of
twenty days was obtained, but the length of survival was not
increased.
In several of the experiments of the series D and E the animals
died without the trypanosomes having reappeared in the blood.
The experiments in series F were undertaken in order to test the
possibility of producing a K 3 -resistant strain of the trypanosome.
Two rats were inoculated with blood from an animal in which the
parasites had first disappeared after the injection of K 3 , and after-
wards reappeared. One of the animals was injected twice with K 3 ,
and the effect of the drug was considerably less than in the other
experiments.
The results here reported are interesting from a general point of
view, and leave no possible doubt as to the powerful action of
salvarsan-copper upon this strain of trypanosome, but they are not
conclusive with regard to the therapeutical value of the drug. After
having concluded my experiments, I became acquainted with the
paper by Van den Branden (1913) on the effect of K 3 in human
trypanosomiasis^ His results are far superior to those obtained by
me in rats and with a different species of Trypanosoma ; this confirms
the information given me by Professor Ehrlich that K 3 has a
markedly different effect upon different trypanosomes.
The experiments here recorded were carried out whilst I was
working for the Yellow Fever (West Africa) Commission of the
Colonial Office, at the Medical Research Institute, at Yaba, near
Lagos, Nigeria. To the Director of the Institute, Dr. A. Connal,
I am indebted for the facilities afforded me.
REFERENCES
Branden, F. van den (1913). Note preliminaire aur quelques e*sais de traitement de U
Trypanose Humaine par Salvarsankupfer. Arch . /. Scb . u. Tropen-Hyg ., Leipzig,
Dec., XVII, 14, pp. 845-849.
Pettit, A. (R913)* Proc6d6 simple pour pr£lever du sang che* les petit* rongeurs.
C. R . Soc . Biol ., Paris, 10 jan., LXXIV, 1, pp. 11-12.
Result
Date t
Rat inoculi
No. tion.
7 'J
l6
*7
1
18
*9
20
161
22.X
... j
!
f
... ,
Death 13 days
. 1
after inoculation 1
Spleen very much enlarged
162
22. X
..... 1 1
... ,
1 1
n
7
»
-
Peritoneal infection
* 6 3
22.X *
1
- | „
*4
ff
n
164
22.X -
i ff
1
8
ff
tt
Spleen very much enlarged
169
22.X
1
ff
11
ff
tt
tt »
165
22.X -
ft
*3
ff
tt
tt tt
166
22.X -
...
ff
H
ff
tt
tt tt ;
167
22.X
...
ff
*7
ft
tt
tt tt
168
22.X
ff
*7
tt
tt
tt tt
1 59
22.X
ff
7
tt
tt
a a
160
22.X -
ff
*4
tt
tt
tt tt
1 95
.5x1 -
...
...
...
1
Death
12 days after inoculation
196
15-Xl
*3
tt
tt
*97
15.XI
13
tt
tt
198
1 1 5.XI + + +
-f 4 ' 4 *
ff
29
tf
tt
) 1S-XI+ + +
+ 4 -+
ff
28
tt
tt
t 9 J
1 ' 5 X 1
\ ff
*5
ff
tt
220
28.XI •••
...
Death 28 days after inoculation
221
28 .xi -
...
a 23
229
I.X1 -
...
...
: l
tt 22
23O
2.X! -
1
... | ...
1
n 32 tt a
23 *
. 3 -xi - j -
j
tt 33
232
4 * - ; -
» 22 „
[p.T.O
240
8.XII
1
13.XII
...
... i
Death 11 days after inoculation
I
2 4*
8.XII
13.XII
2 ^
Necrosis at the site of injection
2 43
10.XII
...
»» *7 11 »»
Large necrotic cavity at the site
of injection
2 44
10.XII
13XH
*
...
ji 3 11
2 47
n.XII
13.XII
...
4 n »>
2 5°
12.XII
13XII
1
1
1 _
10 ?»
Spleen very slightly enlarged
3o;XII
4.I.14
...
...
Death 19 days after inoculation
3a.XlI
4 I «4
1
1
inj. of
0-0064 g.
n 2 ^ » »»
#
?appearcd ; subsequent treatment with K 8 .)
22 7
i.XII
4.XII
- 4 -
+ + +
+ + +
4 - 4 - +
Death 25 days after inoculation
228
1.XII
...
*3 )» »>
201
STUDIES IN BLACKWATER FEVER
III.—THE RELATIONSHIP OF QUININE
TO BLACKWATER FEVER*
BY
J. W. W. STEPHENS
SIR ALFRED JONES PROFESSOR OF TROPICAL MEDICINE, UNIVERSITY OF LIVERPOOL
AND
W. STOTT
HON. STATISTICIAN iO THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
(Received for publication 24 February , 1915)
In the literature of blackwater fever contradictory statements are
made concerning the role of quinine, some observers asserting that
quinine induces the attack of blackwater, others denying this. The
solution of the question is not advanced by the affirmation or denial
of supposed authorities. What we have tried to find out is whether
a statistical examination of the times at which quinine is given
and blackwater occurs respectively, reveals any relationship between
the two.
In examining the literature, an initial difficulty confronted us,
viz., the incompleteness of the records. For our purpose it was,
as will be seen, necessary to know as exactly as possible at what
hour quinine was administered and at what hour the black¬
water ensued. It was often impossible to ascertain these facts,
and we would state clearly that the only data we have used are those
in which the exact times of the giving of quinine and the occurrence
of the rigor or blackwater are stated.
We have thus not considered cases where the statements are not
more definite than, e.g., ‘quinine was given the day before,* ‘the
day of,* ‘shortly before the blackwater,* etc., etc., and also, of
course, those cases could not be considered where it was stated that
no quinine had been taken.
As it was practically impossible to get records of cases with
• Part I. Annals of Trop. Med . and Parasit. (1913). Vol. VII, p. 479,
Part II. Ibid. (1914). Vol. VIII, p. 639.
202
a complete quinine history we were not able to study such,
consequently we proceeded to examine whether any relationship
exists between the time of the blackwater and the time of the
last dose of quinine.
In some cases the time of the rigor is stated, in others the time
of the blackwater. Very probably the time of the onset of the rigor
marks the real onset of the attack and is perhaps more accurate in
that respect than the time of the passage of blackwater, for we have
no accurate knowledge as to how long haemoglobinous urine can be
retained, although it seems to be generally assumed that haemo¬
globinous urine is irritating to the bladder.
The data then that concern us are (i) the time when the last dose
of quinine was given; (2) the time of onset of the rigor; (3) the time
of first passage of blackwater.
We have obtained these or some of these data in 372* cases, the
records of which we have examined.
Time of quinine: This was given in 157 out of 372 cases.
Time of rigor: This was given in 121 out of 372 cases.
Time of blackwater: This was given in 217 out of 372 cases.
It will be noted that it is possible either (1) that the 157 cases in
which the time of quinine is given are none of them the same cases as
the 121 cases in which the time of the rigor is given, or (2) that the
157 cases (except 2) of quinine are none of them the same as the 217
cases in which the time of the blackwater is given, but although both
these possibilities could not hold good for all the cases in categories
(1) and (2), they might do so for a part of both, so that in the
first table the fields of observation are not (entirely) coincident. At
present we disregard this.
The data were now arranged as in Table I. We have taken
eight three-hour periods, because it was (frequently) impossible to
define the time more accurately, and also because if the day were
divided up into twenty-four hours the number of cases falling under
each hour would be too small whereon to base any deductions.
On examining the columns of deviation from an average, it will
be noticed that each has a very decided maximum.
(1) Time of taking quinine. Upon the assumption that a random
_ \ _
• Four cases have been excluded in which the interval between the quinine and the onset of
the rigor or blackwater was greater than 24 hours.
203
distribution* is a priori to be expected, it can be calculated that the
large deviation, 28 in excess of the average, would only be expected
to occur once in over 10,000 cases, so we may fairly say
that the distribution is not a random one, and that there is some
reason for taking quinine between the hours of 6 a.m. and 9 a.m.
This we can assume is due to our methods of life, the time of
rising in the morning determining the time of taking quinine.
Table I
Hour
Number
of people
taking
quinine at
stated hour
Deviations
from
average of
20 per 3
hours
Number of
rigors
occurring at
stated hour
Deviations
from
average of
15 P cr 3
hours
Number of
cases of
blackwater
at
stated hour
Deviations
from
average of
28 per 3
hours
1
12— 3
3
-17
8
- 7
,8
—10
3-6
18
— 2
7
- 8
15
-«3
a.m. J
!
6— 9
48
+ 28
14
— 1
20
- 8
,12— 6
26
i + 6
38
+ 23
47
+ 19
> 2—3
12
- 8
1
22 |
+ 7
5 °
+ 22
3-6
22
+ 2
10
~ 5
37
+ 9
P.M. -
6-9
16
! -♦
9
- 6
17
-11
1 9 — 12
12
- 8
>3
— 2
1
x 3
-■5
.* j
121
1
2I 7
1
(2) The onset of rigor . The maximum deviation in this case is
between the hours of 9 a.m. and 12 noon, Here we have 23 above
the average, and it is also at least 10,000 to one that this does
not occur at random, that is, there is some cause at work which
determines the time.
(3) Passing of blackwater . Here we have the maximum, 22,
• By a random distribution in this case, we mean the distribution that would be expected to
occur, if each case was entered at random in any of the eight divisions into which the day was
divided, by a person who had no bias in favour of one hour rather than another, or if the division
into which each case was entered was determined by picking a ball out of a bag containing eight
differently coloured balls. Out of 157 cases we should expect about 20 in each of the eight
divisions, but almost certainly there would be slight departures, owing to our random method of
filling the divisions, but we have in our 6-9 a.m. division as many as 48 that is a departure from
the average, 20, of 140 per cent.
204
between 12 and 3 p.m., but there is also a large positive deviation,
19, between 9 a.m. and 12 noon. This would be expected to happen
if the period between quinine and blackwater were a little over
3 hours, many cases falling on the border line at noon. Again such
a large deviation as 22 would only occur by chance once in 10,000
cases.
If now we move the figures for the rigor 3 hours back, the
reason for so doing we shall presently see, and compare them with
those of the quinine, we find that the coefficient of correlation ( r )
between them is r — 92 + *04, a very strong correlation (r = 1
is perfect correlation). In non-mathematical language, correlation,
positive or negative, means 1 fit/ and we may illustrate firstly the
meaning of positive correlation in the following way: —
Take sheets of paper in pairs, subject each pair to some operation,
e.g., cutting with a pair of scissors through both at the same time;
if we cut all by a straight line, it would be impossible, when the
sheets were scattered, to bring the pairs together again, because
many pairs would fit one another, but if we made a random
zigzag cut through each pair, then it would be possible after
separation to bring the pairs together, and the certainty we should
feel that we had obtained a pair cut at the same time, would be so
much the greater, the greater the number of zigzags and the greater
their distinctive peculiarity (fig. 1).
To illustrate secondly the meaning of negative correlation, we
may similarly imagine a number of single sheets of paper cut across
by a zigzag cut. When a fit is obtained between two pieces
(belonging to an original whole) it is now a negative one, i.e., the
peaks and depressions of one piece fit the depressions and peaks of
the other. And here again the greater the number of peaks
and depressions that fitted the greater the certainty that they
belonged to an original whole (fig. 1).
Now the method used in calculating the correlation coefficient
gives due weight, both to the number of zigzags, and to the
magnitude of the peaks or depressions. The symbol used for the
correlation coefficient is r, and its numerical value varies from
— 1 to + 1. r = o means no fit, r = + 1 means perfect positive
fit, 7 = - 1 means perfect negative fit.
Except in artificially chosen examples, we cannot expect perfect
I. To illustrate correlation or fit.—There is a negative fit between the two pieces of
paper A and A.i.—i.e., when A is inverted, as in the figure, the depressions of A fit
the peaks of A.i and the peaks of A fit the depressions of A.i. There is a positive
fit between the two pieces of paper I and 2—i.e., the peaks of I fit the peaks of 2,
and the depressions of I fit the depressions of 2 .
206
fit, because we always have errors of observation, and errors due to
random sampling, but the nearer we get to i, positive or negative,
the greater our belief in a connection.
Now the fit in the figures for quinine and rigor is very close,
.92 + ’04 (instead of 1). As we said before, in obtaining a positive
fit between two pieces of notepaper we should feel the more certain
that they were cut at the same time the greater were the number of
zigzags (and the greater their distinctive peculiarity). So that if 24
zigzags fitted, the certainty would be greater than if there were only
12 zigzags. Our cases, however, are insufficient to allow of our
distributing the quinine and rigor figures over every hour of the
day, i.e., into 24 observations, which would be desirable.
We have said above that the fields of observation in Table I
were not coincident, we now proceed to consider those cases only
in which the fields are coincident, i.e., in each case the conditions
affecting the taking of the quinine and the onset of blackwater are,
so far as we can tell, identical.
(1) We have 88 cases in which both the time of quinine and the
time of rigor are given.
Tabli II
Number of
people taking
Deviations from
Number of rigors
Deviations from
Hour
quinine at
stated hour
average of
11 per 3 hours
occurring at
stated hour
average of
11 per 3 hours
|
12— 3
2
- 9
7
“ 4
3— 6
IO
:
! 3
- 8
A.M. ^
|
6— g
3 °
+ 19
9
- 2
' 9—12
1 21
+ IO
!
33
+22
t 12 3
2
- 9 1
15
+ 4
3— 6
11
0
6
- 5
P.M. -
®— 9
6
: “ 5 1
7
~ 4
' 9—12
i
6
1 " 5 !
8
- 3
1 88
i 1
’ 88
i
The correlation between these deviations as they stand is r = 34,
but when the figures for the rigor are moved three hours back the
correlation is r = *85.
207
IZ -3 3-6 6- 9 9-/2 JZ' 3 3-6 6-9 9 ~/z
F Lee so n. del
Chart I. Showing the relationship of quinine and rigor in 88 cases. The mode
of quinine is at 6—9 a.m., that of the rigors is at 9 a.m.—12 noon.
208
(2) We have 103 cases in which both the time of quinine and the
time of blackwater are given.
Table III
Hour
Number of
people taking
quinine at
stated hour.
Deviations from
average of
13 per 3 hours
Number of cases j
of blackwater
occurring at
stated hour
Deviations from
average of
13 per 3 hours
1 l2_ ~ 3
1
—12
4
“ 9
A.M. -j
; 3-6
12
— 1
3
—10
1
j
6- 9
35
+22
9
- 4
1
L 9 — 12
26
+ 13
30
+ 17
1
l ^ 3
4
- 7
21
+ 8
1
1 3- 6
*3
0
22
+ 9
P.M. J.
J 6 — 9
8
- 5
7
- 6
l 9—12
4
“ 9
7
- 6
103
i °3
The coefficient of correlation of the deviations as they stand is
7 = ‘34, but when the figures for the blackwater are shifted three
hours back the correlation is increased to r = '95 (perfect correla¬
tion, 7 = 1).
The increase in the correlations from 34 to ‘85 in one case and
from *34 to ‘95 in the other, when the figures for the effect (rigor or
blackwater) are moved through three hours to fit the (presumed)
cause (the quinine) is very significant, and with such a high correla¬
tion, ‘85 in one case and 95 in the other, it is highly probable that
there is a connection between the quinine and rigor or the quinine
and blackwater, but it is still possible that there is none.
For it must not be assumed that perfect correlation means that
we have obtained a p 7 oof that the events are connected as cause and
effect.
(1) It is possible, for all we know to the contrary, that the mode*
# By a mode is meant the most frequent occurrence of any particular fact or event—e.g., if
the ages of six persons are 5, 5, 5, 5, 5 and 35 respectively, the average age is 10, but the
mode is 5.
209
/ 2-3 3-6 6-9 9 - /2 / 2-3 3-6 6-9 9-/2
P-Le fSON.^cl.
Chart II. Showing the relationship of quinine and blackwater in 103 cases. The
mode of quinine is at 6—9 a.m., that of blackwater is at 9 a.m.—12 noon.
210
(i.e., the most usual time) of the rigors in blackwater, from the
nature of the disease, t occurs at 9 a.m.-i2 noon. If this be so, then
of course a strong correlation might exist between it and any other
(antecedent) phenomenon that had a mode at some antecedent time.
For example, if we investigated the time of taking coffee we
should probably find that the mode (i.e., the most usual time) was
between 6 a.m. and 9 a.m., and so by moving the figures for the
blackwater three hours back we should get a very strong correlation.
It is necessary, then, to alter the conditions, either by altering the
time at which quinine is taken or by observing a sufficient number
of cases when quinine was taken in the afternoon, i.e., in this
particular instance where there is no mode for coffee. If, then, we
find that any alteration in the time of taking of quinine is followed
by a similar alteration in the time of the rigor, the proof of
connection is correspondingly strengthened.
(2) It is possible that the modes which we have found do exist
for quinine and rigor or blackwater, although they exhibit a high
correlation, yet are not really related to one another as cause and
effect. Both modes might be due to some other cause or causes; for
instance, the rise in the price of wheat and the rise in the male death
rate (between which there might be a strong correlation) might be
due to the war, and yet we could not argue that the rise in the price
of wheat was the cause of the rise in the male death rate.
To sum up, we have really started with the assumption that the
taking of quinine by persons with a certain diathesis is the cause,
or a cause, of ‘blackwater.’ On this assumption we should expect
to find a positive correlation between the time of taking the quinine
and the symptoms of the illness.
We do find such a positive correlation, and as the effect follows
the cause usually after a certain period of time, we should expect the
correlation to be brought nearer to unity as the figures for the effect
are brought more in coincidence with the cause, in technical terms
t We find, for instance, the following statements with regard to malaria in Mannaberg (1905):—
‘ Maillot and the majority of observers since him state most paroxysms (about two-thirds of all
cases occur, between midnight and midday—in other words, in the morning/ Moreover, according
to Maillot, 4 the greatest number of quotidian parasites occur about 10 a.m. ; the smallest number
between 9 p.m. and midnight/ Again, it is stated that * Maurel observed in Guiana the majority
of paroxysms between 2 and 5 p.m/ Mannaberg himself states 4 that out of 107 cases 91 per cent,
of the paroxysms occur during the period between 10 a.m. and 3 p.m/ We have not so far been
able to examine these statements critically, so can form no estimate as to what they are worth.
211
as we bring the two modes together. This also we find. Had the
quinine prevented blackwater we should have expected a large
negative coefficient, and if the quinine had no effect we should have
expected the correlation to be practically nothing.
From what we have said, there are then three possible explana¬
tions of this correlation which we have found to exist between quinine
and blackwater.
(1) It is purely accidental, due to the fact that blackwater has a
‘natural* mode in the 9 a.m.-i2 noon interval. This, as we have
said, could be eliminated by seeing whether the same mode existed
when quinine was taken at other times than 6-9 a.m. For this
purpose we require further cases with the times accurately recorded.
(2) The correlation is produced by some unknown cause or causes
which determine both the fact that the mode of quinine is at 6-9 a.m.,
and that the mode of the rigors is at 9 a.m.-12 noon. We cannot
suggest any such cause.
(3) That the correlation is one of cause and effect.
We believe, then, that, provided (2) can be excluded, that an
examination of further cases, in the way we have done here, will
decide whether (1) or (3) is the true answer to the question.
By applying the statistical method adopted in this paper, we
thought it possible that a solution of the question might be
forthcoming, and although the cases we have been able to obtain are
neither sufficiently numerous, nor recorded with the necessary
accuracy of detail for us to claim that a solution has been arrived at,
yet we believe that as a result of our analysis the problem can now
be approached, when further cases are forthcoming, in such a way
that we may fairly anticipate a solution in the near future.
That this is a matter of vital importance, and not simply one of
scientific interest, everyone who has any knowledge of the disease
must realize.
REFERENCES
Broden, A. (1906). L’hemoglobinuric an Congo. Trav. Lab. Med. Leopoldville, Bruxelles,
II, pp. 1-70.
Campenhout and Dryepondt (1901). Fiivre bilie use hemoglobin urique. Trav. Lab. Med.
Leopoldville, Bruxelles, I (1899-1900), pp. 51-117.
Christophers, S. R., and Bentley (1918). Blackwater Fever. Scientific Memoirs by Off.
Med. San. Dept. Govt. India, Simla, No. 35.
Decks, W. E., and James, W. M. (1911). A report on hemoglobinuric fever in the Canal Zone.
A study of its etiology and treatment. Pp. 177. With charts and tables. Mount
Hope, C.Z.
Doering (1898). Ein beitrag cur kenntniss der Kamerun-malaria u.s.w. Arb. a. d. k. Gesund-
heitsamte, Berlin, XIV, S. 121 -137.
Kleine, F. K. (1901). Ueber Schwarxwasserfieber. Zcitschr. Hyg. Inf., Leipzig, XXXVIII, pp.
472-486. With charts.
Koch, R. (1899). Ueber Schwarzwasserfieber (Ilamoglobinurie). Zeitschr. Hyg. Inf., Leipzig,
XXX, pp. 295-327.
Maillot (1836). .' Traite des fievres intermittentcs. Paris, 1836.
Mannabkrg, J. (1905). Malarial Diseases. Nothnagel’s Encvcl. of Practical Medicine.
English Edition, W. B. Saunders & Co., Philadelphia and London, p. 226.
Maurel (1883). Traite des maladies paludiennes. Paris, 1883.
Pampoukis, P. S., and Chomatianos, S. (1889). Recherches cliniques et experimentales sur
Ph&noglobinurie quinique. Progria medical, Paris, VIII, 2 seric, pp. 3-6.
Panse, O. (1902). Schwarzwasserfieber. Zeitschr. Hyg. Inf., Leipzig, XLII, 1, pp. 1-44.
With charts.
Plehn, A. (1896). Beitrage zur Kenntniss von Verlauf und Behandlung der tropischen malaria
in Kamerun, Berlin. August Hirschwald.
Plehn, A. (1914). Ein beitrag zur kenntniss der akut hamolytischen malaria (Schwarzwasscr
fieber). Deutsch. Med. Woch., XL, pp. 1414-1416.
Plehn, F. (1898). Die Kamerun-Kiiste. Studien zur Klimatologie, Physiologic und Pathologic
in den Tropcn. Mit 47 Abbildungen und 1 Karte. Berlin, A. Hirschwald.
Reports (1912). Reports on Blackwater Fever in the Tropical African Dependencies.
Reports (1914). Blackwater in the Tropical African Dependencies. Reports for 1912.
Schellong, O. (189 a). Die Malaria Krankheiten u.s.w. Berlin, 1890.
Stephens, J. W. W., and Christophers, S. R. Reports to Malaria Commission of Royal Society,
London. (1900) 1st Ser., pp. 12-42, pp. 42-75 ; (1901) 5th Ser., pp. 12-27; (1903)
8th Ser., pp. 3-26.
Tomaselli, S. (1897). La Intossicazionc clinica e. l’infezione malarica, Catania, pp.
XVI +- 204.
Volume IX
June, 1915
No. 2
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. VV. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor WARRINGTON YORKE, M.D.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool .
C. Tinting & Co., Ltd.
Printers to the University Press of Liverpool
53 Victoria Street
21 $
GANGOSA IN NEW GUINEA AND ITS
ETIOLOGY
BY
ANTON BREINL
FROM THE AUITRAL1AN INSTITUTE OF TROPICAL MEDICINE
(Received for publication 20 May , 1914)
Plates XIV-XVII
Gangosa, or Rhinopharyngitis mutilans, is a disease of limited
geographical distribution, being, according to our present know¬
ledge, confined mainly to the Pacific Islands.
According to Castellani and Chalmers (1910), cases of this
disease were described first in 1828 by a Spanish Commission to the
Marianne or Ladrone Islands, under the name of Gangosa,
meaning ‘Nasal Voice.* Fordyce and Arnold (1906), and in the
same year Leys, gave a clinical account of a similar disease in Guam,
and were followed by Mink and McLean (1906) who discovered cases
of Gangosa in the Ladrone and Caroline Islands.
, . Later, Stitt (1907) reported the occurrence of Gangosa in a white
man, who was treated in the United States Naval Hospital at
Canacao. This patient had contracted his infection during a stay
in Guam, where he had become intimately associated with natives
in whose families Gangosa had been known.
A further case with complete post-mortem findings was
described by Musgrave and Marshall (1907) in a native of Santo
Domingo de Basco, Batan Islands, being the first case reported
from the Philippines. Careful examination of the blood was made
by these authors. According to their findings the blood did not
show any marked pathological changes; the differential count
proved that the relative numbers of the various types of leucocytes
were normal.
In 1909, N. T. McLean observed two patients suffering
from what he believed to be Gangosa on the occasion of a visit
214
to the Hospital at Gonaives, Haiti, and stated that according to
information that he collected numbers of such cases were supposed
to have occurred at Port au Prince.
A case was also published by Stitt in a young Filipino who had
never been outside Cavite Province (Philippine Islands).
In 1910 Garrison pointed out that there were 327 cases of
Gangosa in Guam, and pronounced the opinion that according to
his experiences from a clinical, and particularly from a
therapeutical standpoint, Gangosa was to be considered a very
late form of Syphilis—perhaps a fourth stage. Odell (1911) reported
a positive Wassermann reaction in 82 % of his cases. Anti¬
syphilitic treatment seemed to benefit all cases considerably.
Rossiter (1912) described a disease in a two year old child where
the destruction of the hard and soft palate were the most prominent
lesions. In specimens taken from the ulcerated surfaces numerous
Spirochaeta per tenuis were found. Two months previously the
child had suffered from typical framboesia.
Also in 1912 Ziemann drew attention to the fact that a disease
resembling Rhinopharyngitis mutilans occurs not infrequently
amongst the negroes of Kamerun.
I observed cases of Gangosa for the first time in the Torres
Straits Islands, whilst on a short journey during the latter end of
1910.
Then I was inclined to consider this disease either as a peculiar
manifestation of syphilis or as a hitherto undescribed disease
occuring on Murray Island. Elkington referred to most of these
cases in the yearly report of 1912, adding a number of cases which
he discovered on Darnley, Boigu, Saibai and Dauren Islands. The
disease must have been prevalent on Murray Island for nearly a
century, as it is, according to Elkington, referred to by Dr. Wilson,
R.N., in an account of a visit made by him to Murray Island in
1822.
The occurrence of cases of Gangosa in New Guinea has been
noted previously, but these cases had been regarded by Sir William
Macgregor as lupus.
During my short journey in New Guinea typical cases of
Gangosa were seen in villages on the south coast, especially in
Kerapuna, a fairly large village, where as many as ten cases were
215
examined. Furthermore a number of natives suffering from
Gangosa were encountered in the western parts of British New
Guinea and in the Mekeo District, and there the earliest stages of
the disease were seen, showing peculiar manifestations which did
not altogether coincide with the description given by previous
observers elsewhere.
CLINICAL ACCOUNT OF THE DISEASE
According to Castellani and Chalmers the disease ‘ begins as a
sore throat, when on examination a nodule can be seen on the
back of the pharynx, the posterior pillars of the fauces or the
edge of the soft palate. This ulcer usually spreads rapidly at first,
but more slowly later, and destroys the soft palate first, later the
bony parts of the palate and the nasal septum. In consequence the
skin of the nose falls in, and the cavity of the nose and mouth are
connected.
1 In consequence of the destruction of the soft and hard palates
the ulceration may extend afterwards to the face or lips or affect
the pharynx. As a rule a pungent odour is given off, whilst a
slight discharge of granular and necrotic debris is present. The
ulceration may progress continuously for a period of 10-35 years,
or it may advance at certain times and be quiescent at others, or it
may cease at any time, leaving a chronic ulcer.*
So far as observed, the cases of Gangosa met with in New
Guinea may be conveniently divided into three groups.
Firstly. Early cases in which the patients had been ill for a
few months only and where a small area of the skin of the face was
found to be affected. The cartilaginous septum had been destroyed
but the disease had not caused any extensive destruction of the
bone.
Secondly . Cases in which the ulceration had advanced
further. Large areas of the face were found to be the seat of
extensive weeping sores. The bone near the affected parts had
become implicated, and the disease had led to the destruction of
the skeleton of the nose and the hard palate.
Thirdly. Cases in which the active ulcerating process had come
to a standstill. Extensive cicatrisation had taken place at the
21 6
seat of the ulceration, as a rule the nose had disappeared, and was
represented by a smaller or larger opening. In one extreme case
the opening was so small that the woman in question was able to
smoke comfortably a cigarette through the small opening—the only
remains of the nose. In another case the whole of the back of
the pharynx could be seen without any difficulty through the large
opening in the middle of the face.
The uncertain etiology of Gangosa and its similarity to tertiary
syphilis in its clinical manifestations makes it advisable to refer
in greater detail to the history of a few of the typical cases met with.
EARLY CASES
Only a comparatively small number of early cases were seen.
Case i. (PI. XIV, fig. i.) Apau Kaianga was an unmarried girl of about
12 years of age. Her parents as well as four brothers and sisters were alive and
well. Her disease began about one month previous to my visit as a sore below
her nose which spread rapidly, affecting especially the septum, the left ala of the
nose, the upper lip and the left corner of the mouth, and had already destroyed
part of the cartilaginous septum.
The examination revealed the nose considerably flattened, and also thickened on
account of oedematous swelling. The left ala was the seat of a large raised dry ulcer,
covered by a thin scab, below which there was a reddish uneven granulating surface.
This ulceration had extended to the upper lip, being confined to its central part,
just below the nostrils. The left corner of the mouth was the seat of a similar
ulcer which showed an uneven surface. The upper lip around the ulceration
was swollen and oedematous and its surface was intersected by purplish irregular
raised lines simulating scar tissue. The cartilaginous septum had been partly
destroyed by the ulceration, whereas the osseous septum .was still intact. The
hard and soft palate were of normal appearance.
Case 2 (PI. XIV, fig. 2.) Ame Aua was a woman about 25 years of age,
belonging to the same village. Her father was still living, her mother had died.
Some time ago she was married, her three children were alive and well, except
for manifestations of Yaws.
According to her own statement she had been ill for four months only, the
disease nn her had begun as a small sore on the tip of her nose which had spread
gradually into the nasal cavity. The examination revealed a fairly large ulcer
on the nose, a yellowish greasy scab covering the raw granulating surface. The
cartilaginous septum of the nose had been destroyed by ulceration and the nose
was, in consequence, flattened. A similar ulcer below the nostrils extended into
the nasal cavity. On the right cheek was a narrow raised line resembling oedematous
scar tissue. The hard and the soft palate and the pharynx did not show any
pathological changes.
2I 7
Case 3. (PI. XIV, fig. 3.) The woman belonged to Kerapuna, a large village
on the south-east coast of New Guinea. She had been married and was the mother
of six children. Four of them were healthy and two were in the secondary stage
of Yaws.
Her upper lip showed the same scar formation as described in the two previous
cases, the scars being oedematous and raised in the centre. The free edge of the
skin of both the alae showed ulceration and both nostrils were closed up by yellowish
scabs.
Case 4. (PI. XIV, fig. 4.) Ame Kwipa, a young girl, belonged to Veipa, a
large village in the Mekeo District. She had had small sores on her face, especially
on her right cheek for about one year, and these had shown a tendency to heal
during the last two months. Her brothers and sisters were healthy.
When examined her nose appeared broadened, its skin swollen and oedematous.
The upper lip, especially the right side, was swollen, oedematous, and showed
raised irregular lines resembling scars. The right cheek was the seat of a
number of small pustules which contained whitish pus.
Case 5. (PI. XV, fig. 5.) Ainye, a girl about 14 years of age, belonged to
Bebeo, a small village in the Mekeo District. Her father was alive and well,
her mother and one brother had died. Her illness commenced about one year
previously with a small sore on the upper lip below the nose, which had spread
slowly and gradually to the nose and downwards to the mouth.
When examined her nose was flattened, showing an uneven surface; the mucous
membrane of her nostrils was covered by a yellowish scab. The upper lip was
swollen and succulent in appearance, whilst the left angle of her mouth was drawn
up by the formation of scar tissue. Her body was clean.
Case 6. (PI. XV, fig. 6.) The patient was a young unmarried girl, Wabnagi,
belonging to Kaile, a village on the south-east coast of New Guinea. Her mother
was alive, her father dead. Her disease began about two years previously as a
small sore on the upper lip just below the nose, which had spread gradually to the
cheeks and into the interior of her nose.
At the time of examination her nose was flattened and had sunk in, as the
cartilaginous septum had completely disappeared. The surface of the nose was
uneven and rough ; the right half of the upper lip was drawn up on account of
extensive scar formation. The same scar formation was seen on both cheeks,
more extensively, however, on the right side. The skin of the affected parts
was very thin and shiny, intersected by raised streaks of apparently dense scar
tissue.. Here and there on the nose and cheeks were small ulcerations covered
by a yellowish scab. The hard and soft palate did not show any pathological
changes.
Case 7. (Pl. XV, fig. 7.) Was a young unmarried woman belonging to
Kerapuna. Her parents as well as two younger sisters were alive and well. Her
disease began when a small child with sores on her face which had since spread,
implicating the nose. She showed large and extensive ulceration on her face ;
the nose had completely disappeared and was represented by a small opening
in the middle of the face surrounded by irregular ulcers covered by yellowish
scabs. The upper lip was partly destroyed by extensive ulceration. A large
ulcer on her left cheek had a raised granulating surface and irregular edges, secreting
copiously a slightly yellowish clear, foul-smelling, discharge. The left cheek
showed scar formation. The hard and soft palate were normal.
218
LATER CASES
Case 8. The patient, a man about 22 years of age, belonged to Hula, a
village on the south-east coast of New Guinea. His mother had died, his father
was alive and healthy. He was married and had three normal children. According
to information the disease had commenced about five years previously with a sore
on the forehead over his left eye-brow, which had spread gradually until the
greater part of his face was affected. His face, when examined, was badly disfigured;
the nose was flattened and was sunk in. Deep scar formation on the right side
of his forehead caused the upper eyelid to be drawn up. The skin on the left
side of his forehead showed extensive ulceration which had spread downwards
and caused the destruction of his left eye. The central part of Jiis upper lip was
partly destroyed. The hard palate was perforated, showing a round hole of
about 2 cm. in diameter, with smooth rounded edges.
Case 9. Was a woman, Kila Alukma, of about 40 years of age, living in the
same village as the previous case. Her parents were dead, her two daughters
were married and apparently well.
Her disease had begun about ten years previously ‘ with a pain in the stomach
which gradually worked up to her mouth and came out through her nose/ She
pointed out that there had been a sore in her mouth before her face became
affected. This case was very similar in appearance to Case 8. There was extensive
scar formation on the face round the nose, the upper lip was drawn up on account
of dense scar tissue. A stinking discharge from the nostrils was noticed; the
hard palate was perforated, the oral and nasal cavity communicating through a
round opening of about 3 cm. in diameter.
Case 10. Was an unmarried woman of about 35 years of age belonging
to Kaile on the south-east coast of New Guinea. Her mother was still alive,
her father was dead. About ten years previously a small ulcer had started on
her nose and soon began to spread. At the same time ulcers appeared on her legs.
When examined the cartilaginous septum of her nose had disappeared and
extensive scar tissue formation was observed around the nose, on both cheeks
and on the upper lip, the mouth and eyelids being normal. The hard palate
was perforated. Her legs, as well as her left forearm were the seat of deep ulcers
of varying size some of which extended as far as the bone, whilst some of them
were covered by a greyish greasy scab.
Case i i. (PI. XV, fig. 8.) Was a man, Aiaba, about 40 years of age belonging
to Kivori village. Both his parents were dead. He was married and the father
of six healthy children. His illness had begun about four years previously with
a small sore on the upper lip just below the nostrils, which spread at first into the
nasal cavity and later to his face. He had lost the toes of his right foot in an
accident when he was a small boy.
The patient’s face was of horrible appearance. The nose had completely
disappeared, leaving a large hole in his face through which the posterior wall of
the pharynx could be seen. A large irregular granulating sore on his forehead
was covered by a yellowish dry scab under which thick pus had collected. A similar
sore was on his right cheek showing here and there recent scar formation. The
upper lip was the seat of extensive scar formation. The skin of his left cheek
was swollen and oedematous.
The hard palate was perforated and the oral cavity communicated with the
nasal cavity by means of a large round opening with sharp edges.
219
THIRD STAGE OF THE DISEASE
Case 12. (PI. XVI, fig. 9.) A woman, Ugre, about 25 years of age, who
lived in Kerapuna, a village on the south-east coast of New Guinea. She had
been ill, according to information, for about ten years; her nose had been affected
long before her mouth became involved. When she fell ill her husband married
a second wife. Both were seen and were in excellent health. When examined
the skeleton of her nose had disappeared and there was a hole in the centre of her
face, the surrounding skin as well as the skin of the upper lip and forehead showed
extensive scar formation, indicating that the whole face had been previously
affected by the disease.
Case 13. (Pl. XVI, fig. 10.) An unmarried woman, about 32 years of age,
was seen in the same village as the previous case. Her father had died, her mother
was still alive and in good health. Her disease had started when she
was quite a small child. When examined the bony skeleton of her nose had
completely disappeared, the nose was sunk in so that the nostrils were only
represented by a small opening. Both the upper and the lower lips had disappeared,
the skin of the face directly joining on to the gum, so that the teeth were
unprotected; the skin of her face was one mass of scar tissue, the right lower
eyelid was drawn down by scars giving rise to an ectropion. The left side of
her throat and her left shoulder were of the same appearance as the skin of her
face, showing extensive scar formation. Her soft and hard palate did not show
any pathological change.
Case 14. (PI. XVI, fig. 11.) An unmarried woman of about 35 years of age.
She had been ill for quite a long time, in all probability between ten to fifteen
years. Both her parents were alive and in good health. Her disease began on
the ‘ outside of her nose 9 in the form of a small sore which had spread slowly
implicating the lips and mouth. Some time ago the ulcerated parts had begun
to heal up. Her face was completely disfigured. The whole outer nose had
disappeared as well as the anterior part of her hard palate, so that the oral and
nasal cavities communicated extensively. The posterior part of the hard and
soft palate were still intact. The central part of the upper lip and the lower
lip had disappeared, the outer skin joining immediately on to the mucous membrane
of the gum. There was extensive scar formation on the skin of her face and on the
neck which closely resembled the scars after deep burns.
Case 15. A woman of about 35 years of age belonged to Ififu (Mekeo
District). Her parents were dead, two brothers were alive and well, one of them
showing scar formation around the mouth, most probably the remainder of Yaws.
She was married and mother of a healthy girl. Her illness began when she was
a girl, in the form of a small sore on the upper lip. Subsequent to the infection
of the face a large ulcerating sore developed on the abdomen, around the navel
which, however, soon healed up. Her mouth was small and deformed, the left
angle drawn up by scar tissue. Extensive scar tissue formation was seen on the
upper lip and both cheeks, the skin otherwise being smooth, shiny and very thin.
Case 16. (PI. XVI, fig. 12.) Was an unmarried woman, Kaiyo, about 35 years
of age. Her parents were dead. The disease began at a very early age with
a sore on the left foot soon after she had begun to walk. Later on a small ulcer
appeared on her upper lip below the nostrils which gradually spread, eating away
the whole upper lip and the nose. At the time of examination, her nose as well
as her upper lip had completely disappeared, a small round hole representing
2 20
the nasal opening. Extensive scar formation was noticeable around the mouth
and on the cheeks. Her left eye had been destroyed by the disease. The hard
and soft palate did not show any pathological changes.
Case 17. Was a middle-aged woman. Her parents were dead. She had
four brothers and sisters, two of whom had died. She had never been married.
Her disease began some time ago with an ulceration inside her nostrils. Her
nose had been destroyed by the disease, and the back of her pharynx was visible
through a large triangular opening in the face. The anterior part of the hard
palate was destroyed, but the posterior part was still existent. Her mouth was
deformed, the right angle drawn up by scar tissue.
The case histories recorded above describing a few typical cases
out of the many examined, prove that Gangosa, or Rhino¬
pharyngitis mutilans, is a definite morbid entity, and can be
differentiated from other diseases causing similar lesions, such as
Syphilis, Lupus and Leprosy. It is a very chronic but very rarely
fatal complaint. A great number of the patients recover without
specific treatment, mostly, however, after the morbid process has
brought about extensive destruction of the face.
From the number of cases observed in the different stages,
Gangosa in New Guinea seems to differ in its earliest stages from
the disease described by Castellani and Chalmers.
According to my experience in New Guinea, Gangosa usually
begins as a small ulcer on the upper lip, just below the nose. Only in
a small number of cases did patients state definitely that the palate
was affected before any other part of the face. This small ulcer
soon begins to spread, destroying at first the fleshy parts of the face,
such as the lips, the alae and the skin of the cheeks surrounding
the nose. As a rule, the tissue in the near neighbourhood of the
ulcer is swollen on account of the oedematous infiltration, the surface
skin is glossy and intersected by reddish, raised lines, resembling
in some respects a relief map of mountains.
In the earliest stages the ulcer may spread gradually or some¬
times rapidly, finally implicating the bones of the infected regions,
destroying at first the cartilaginous parts of the nose, and later on
the bony skeleton and the soft and hard palate.
The ulcers are raised, possess irregular edges, and are not well
defined from the surrounding tissues. The surface of the ulcer is
formed of granulating tissue, which secretes a malodorous yellowish
discharge, and is often covered by a dark yellowish scab under¬
neath which pus accumulates. Very soon, however, some of the
221
ulcers show a tendency towards healing. The granulation tissue
begins to discharge less, and after a varying period new smooth
a skin grows over the ulcer from the surroundings, often, however,
"breaking down again, and giving rise to renewed ulceration.
Some time afterwards dense scar tissue is formed, leading to
the deformities of the face so characteristic of the disease.
Now and again only a single primary ulcer is seen, at other
times a number of ulcers appear simultaneously.
The morbid process may come to a standstill at any stage of the
disease, even before it has led to extensive destruction.
The tendency to scar formation is a marked feature of the
disease, and in this respect Gangosa simulates Syphilis, so that in
some of the late cases a differential diagnosis would be practically
impossible.
Some of the patients who showed typical symptoms of Gangosa
in their faces were suffering from large ulcerations on other parts
of the body, especially on the legs. It is impossible to decide
whether these ulcers corresponded' only to Ulcus tropicum, or were
due to the same parasites as the face lesions.
It is interesting to note that the great majority of cases seen
were women, only a comparatively small number of men being
affected.
THE ETIOLOGY OF THE DISEASE
As the etiology of the disease is unknown, material was collected
for microscopic examination. Special attention was paid to the
early lesions with a view of finding a possible parasite in the
oedema fluid. It may well be understood that the examination of
the secretion of open sores of advanced cases did not seem to offer
much opportunity for the forming of a definite opinion; smears
taken from the secretion of open wounds always contain enormous
numbers of bacteria and spirochaetes of varying shapes and sizes,
which are to be considered as secondary infections.
From the earliest cases showing the marked oedematous swelling
of the affected parts as described above, oedema fluid was obtained
by pricking the skin over the lesion with a needle in the same way
as serum would be obtained from a leprous nodule. The specimens
222
were stained with Giemsa's stain. In the oedema fluid of five early
and two later cases, in addition to a small number of red and white
corpuscles, small bodies were found resembling yeast cells. The
majority of them occurred free, only a small number was contained
in leucocytes (PI. XVII, figs. I and 2). These cells showed a marked
polymorphism; some of them were round, others oval or pear-
shaped (comp. figs. 3 and 4), and were seen either singly or in
groups.
The cells measured from 075 fi to 4 /*, and consisted of a
lightly bluish staining cytoplasm showing a typical network
structure, a great number of them contained a distinct and
sharply-defined vacuole (PI. XVII, figs. 5-7) and small roundish
masses which resembled chromatin in their staining reaction. These
chromatin-staining masses occurred either singly, being more or less
sharply defined, roundish or oblong in shape (figs. 7, 8, 13) or as
number of similar bodies enclosed within the cell (figs. 9, 10,
14. 15)-
Budding of the cells in its various stages could be frequently
observed.
In the earliest stages only a small protuberance on the periphery
of the cell was seen, consisting of cytoplasm only (figs. 3 a, 6, 14).
In the more advanced forms this protuberance had increased in
size, and at this stage a small chromatin-like staining mass was
often seen at the base of the bud (figs. 3 b, 16). Sometimes the bud
itself contained one or more chromatin-like staining particles
(figs. 7, 13).
The bud increased in size, and usually became detached before
it reached the same size as the parent cell (figs. 9, 10, 16), and in
this case invariably contained chromatin-like staining particles and
often a distinct unstained vacuolic area (fig. 18).
In many instances only one bud was formed, not rarely, how¬
ever, two, three or even more buds were seen still connected with
the cell (figs. 7, 15).
Forms as seen in fig. 17 were noticed only very rarely. In these
forms the cell consisted of a central portion stained lightly reddish by
Giemsa's method, containing one or more distinct chromatin-like
particles, the whole being surrounded by a thick bluish pellicle.
From their morphological appearance there is no doubt that the
223
cells described are to be considered as parasites belonging to
the genus Cryptococcus of the family Saccharomycetes. The
characteristic features of Cryptococcus is the reproduction by
budding only and the absence of endospores and ascospores.
In these parasites only multiplication by budding was observed,
endospores have never been seen, although carefully sought.
The parasite itself has all the typical features of yeast cells.
Unfortunately circumstances only permitted of the collecting of
dry films, which were stained by Giemsa’s method, and nothing
definite can be said about the nuclear details. When stained by
Gram’s method the cells did not become decolourised.
Parasites of the genus Cryptococcus are well known to produce
granulating sores in man and beast; the best known and most
studied disease due to Cryptococcus is probably epizootic
lymphangitis in horses, occurring in different parts of the world.
Gilchrist and Stokes (1898) describe a similar parasite in man
from a case of chronic ulcerative dermatitis, resembling Lupus, under
the specific name of Cryptococcus dermatitis. Busse (1894) isolated
Cryptococcus hominis from a purulent periostitis of the tibia of a
woman.
On the other hand, yeast cells occur very frequently in open
sores of every description.
These parasites were found in great numbers in the oedema
fluid of the early cases of Gangosa, where open sores had not
formed. Moreover, specimens of the oedema fluid did not contain
any other microorganisms besides the yeast cells and a small
number of red and white blood corpuscles.
In the seven cases from which oedema fluid could be obtained
these parasites were found, most numerously in Case 11, where early
and late manifestations were present at the same time, and in which
the disease seemed to be progressing most rapidly.
The finding of a Cryptococcus in seven cases out of eight
examined, suffering from Rhinopharyngitis mutilans, justifies the
conclusion that one may regard this parasite as the cause of the
disease in question.
The fact that blastomycetes are known to cause similar skin
lesions in man and animals is an additional argument in favour
of the Cryptococcus being the etiological agent of Gangosa. Wc
224
therefore propose for this parasite the specific name of Cryptococcus
mutilans.
Unfortunately, circumstances did not allow attempts at cultiva¬
tion. Animal experiments have been performed at our instigation
by Dr. Elkington whilst visiting Murray Island. Two monkeys
were inoculated on the eyelid and intra-nasally with the secretion
of open sores, but in spite of prolonged observation none of the
animals in question showed any lesions whatsoever.
CONCLUSION
Rhinopharyngitis mutilans is a disease which occurs not
infrequently amongst the native population of the south coast of
British New Guinea, and is a blastomycosis due to Cryptococcus
mutilans , n.sp.
REFERENCES
Breinl (1910). Report of the Australian Institute of Tropical Medicine for the year.
Busse (1894). Ueber parasitare Zelleinschlusse und ihre Ziichtung. Ccntralbl. f. Bakt.
Bd. XVI.
Castellani and Chalmers (1910). Manual of Tropical Medicine.
Elkington (1912). Annual Report of the Commissioner of Public Health, Brisbane.
Fordyce and Arnold (1906). Journal of Contagious Diseases , Vol. XXIV, 1.
Garrison (1910). Gangosa in Guam. Bulletin of the Manila Medical Society , Vol. II,
No. 10.
Gilchrist and Stokes (1898). The presence of an oidium in the tissue of a case of
Pseudolupus vulgaris. J. Hopkins Hosp. Rep ., Vol. VIII, No. 64.
Leys (1906). Journal of Tropical Medicine , Vol. IX, p. 47.
McLean (1909). Gangosa in Haiti. United States Naval Medical Bulletin , Vol. Ill, p. 141.
Mink and McLean (1906). Journal of American Medical Association , Vol. XLVII,
p. 1166.
Musgrave and Marshall (1907). Gangosa in the Philippine Islands. Philippine
Journal of Science, Vol. II, p. 387.
Odell (1911). Gangosa a form of Syphilis. United States Naval Medical Bulletin, Vol. V,
No. 4.
Rossiter (1912). Report of a case resembling Gangosa in which Treponema pertenuis
was present. United States Naval Medical Bulletin , Vol. VI. No. 1.
Stitt (1907). A case of Gangosa in a white man. United States Naval Medical
Bulletin , Vol. I, p. 96.
-11910). A case clinically resembling Rhinopharyngitis mutilans. United States Natal
Medical Bulletin , Vol. IV, No. 4, p. 524.
Wilson (1822). Voyage round the World.
Ziemann (1912). Discussions during the Meeting of the German Tropical Society. Archiv f.
Schiffs - u. Tropenbygiene, Bd. XVI, Beiheft, p. 85.
226
EXPLANATION OF PLATES
Plate XIV
Figs. 1-4. Cases of Gangosa (Rhinopharyngitis mutilans).
.-{finals Trop. Med.
228
Plate XV
Figs. 5-8. Cases of Gangosa (Rhinopharyngitis mutilans)
A finals Prop. Med. & ParasitolVol. IX
PLATE XV
C. Ttitling «S^ Co., Ltd., Imp
130
Plate XVI
Figs. 9-12. Cases of Gangosa (Rhinopharyngitis mutilans).
Plate XVII
All the figures were drawn with a large Abbe drawing apparatus,
using 2 mm. apochromatic oil immersion.
For Figs. 1-2. Compensation ocular 8 was used.
For Figs. 3-18. Compensation ocular 18.
The specimens were fixed in alcohol and stained by Giemsa’s
method.
Figs. 1-2. Intracellular forms of Cryptococcus mutilans.
Figs. 3-4. Groups of parasites.
Figs. 5-7, 9, 10, 12-16, 18. Different stages of budding.
Figs. 8, 11. Large forms.
Fig. 17. Apparently encysted form, rarely seen.
(4 ■
\:
%
■ % .#
f
i.
Vv.’
4
a
4
.7
0
k f
v ;
-**
\
•
• •
••
fc.
,v
fit
f* r * x
/ ^
! - r * A
m
»
It
rj
n
a
0 #*.
•:
* ^
f-- ^
*
/.;
to
17
/<v
(rfut/t/M AW»'/ /s </r/
CRYPTOCOCCUS MUTILANS.
233
THEILERIA TACHTGLOSSI (n.sp.)
A BLOOD PARASITE OF TACHT-
GLOSS US ACULEATUS
BY
HENRY PRIESTLEY, B.Sc., M.B., Ch.M.
FROM THE AUSTRALIAN INSTITUTE OF TROPICAL MEDICINE
(Received for publication 20 May y 1914)
Plate XVIII
Hitherto blood parasites of Monotremes have not been
described.
Gilruth, Sweet and Dodd (1911) refer to the occurrence of
Attaplasma marginalc (Theiler) in the blood of Echidnas, but
according to their conception the presence of these bodies has no
pathological significance.
The blood of an Echidna, from the neighbourhood of
Townsville,* on examination, showed the presence, within the red
blood corpuscles, of numerous parasites resembling the Thcileria of
East Coast Fever. On rare occasions the same structures, referred
to by Gilruth, Sweet and Dodd as Anaplasma } could be seen.
When the blood of the animal was again examined after an
interval of a week, most of the parasites had disappeared from the
peripheral circulation, and so the animal was killed and films made
from the blood and different organs.
Dry films were fixed in alcohol and stained with Giemsa’s
solution. It was found practically impossible to obtain satisfactory
results with wet fixing and staining methods for the blood films. On
the other hand the detailed structure of Koch’s bodies could be
studied to advantage in organ films after wet fixation in sublimate
alcohol and staining by Giemsa’s method.
* I take this opportunity of thanking Mr. J. Humphrey of the Haughton River through
whose kindness the animal was obtained.
234
MORPHOLOGY OF THE PARASITES IN THE PERIPHERAL
BLOOD
In their morphological appearance the parasites resembled
closely Thcileria parva of East Coast Fever.
They occurred, invariably, within the red blood corpuscles
without causing any apparent alteration in the shape and form of
the cells.
Bacilliform, ovoid, pyriform, round and comma-shaped
parasites were seen, but on careful examination intermediate forms
were often encountered. Typical examples of each class could,
however, be easily distinguished.
The bacilliform parasites (PI. XVIII, figs. 1-3) occurred most
frequently. They measured i‘S yu to 3 o yu by 0*3 M to O'8/x, and
were cither perfectly straight or slightly curved.
The cytoplasm stained dark blue and the chromatin was
situated either in the centre or at one end of the parasite.
The comma-shaped parasites (fig. 5) measured 2 0 yu to 3 0 n
by 0 8 yu to 1*2 yu, were broader and were rounded off at one end.
The cytoplasm stained less deeply and an unstained vacuolar area
was frequently present.
The ovoid and pyriform parasites (figs. 4 and 6) measured 1*0 yu
to 3*0 ft by o‘5 yu to 1*2 yu, and no sharp line of distinction could
be drawn between these two types. A vacuole was almost always
present, with the chromatin, as a rule, in close proximity to this
unstained area. Sometimes, however, the chromatin occurred in
the form of an irregular band across the centre of the parasite.
Rounded forms (fig. 7) were the least frequently encountered.
They measured ro yu to 2*2 yu in diameter, and resembled the ring
forms of malarial parasites. The cytoplasm, which stained light
blue, appeared to be condensed peripherally, surrounding a central
clear area. The chromatin occurred as an oval or round mass, or
was crescentic in shape and situated at tne periphery. Sometimes
two masses of chromatin were seen in the same parasite.
Multiple infection of red blood corpuscles was very rare. In
five slides three corpuscles were found containing two parasites,
one contained three parasites and one contained four parasites
(fig- 9 )-
The so-called cross forms with chromatin particles at each pole
were seen on two occasions only (fig. 8).
2 35
As in infections with Theileria parva , the so-called Koch’s
bodies occurred in small numbers in the peripheral blood, but in
great numbers in smears made from the organs, particularly the
liver, spleen and lungs. They were either free or included in
leucocytes or endothelial cells, and represent in all probability an
asexual multiplication of the parasite.
The smallest forms within the leucocytes somewhat resembled
cell granules. They were about i fi in diameter, rounded, with
pale blue cytoplasm and a comparatively large chromatin mass in
the centre. The largest forms occurring intracellular or free in
about the same proportion were rounded or oval in shape,
measuring up to 12 n in diameter.
The cytoplasm stained by Giemsa’s solution was either dark or
light blue, showing the typical network structure; the chromatin
occurred in the form of numerous granules, irregular in shape and
size and unevenly distributed throughout the parasite. In many
instances a great number of chromatin granules were aggregated
into masses forming irregular bands across the cytoplasm, at the
same time chromatin particles were dotted over the remainder of
the parasite (fig. 16).
All intermediate stages in the leucocytes were observed between
the smallest forms containing only one small chromatin granule
and little cytoplasm and the largest forms with a great number of
chromatin granules. The cytoplasm of the largest intracellular
forms was usually well defined from the cytoplasm of the host
cell; in some cases, however, no such differentiation could be made
out, and in these instances irregular chromatin granules were
unevenly distributed throughout the host cell. It would seem that,
in these cases, the parasite had ruptured, leaving the chromatin
granules, most probably surrounded by a very small amount of
cytoplasm, free within the host cell. The fact that, now and
again, fully developed parasites, of the same type as those found
in the red corpuscles, were seen in leucocytes in addition to
chromatin granules, suggests that the majority of the chromatin
granules may develop into the small blood forms.
As in the case of Theileria parva , when the leucocyte contains
a large Koch’s body the cell nucleus may be disintegrated.
The parasite of the Echidna closely resembles, in its life
history, Theileria parva as described by Gonder.
It may be assumed from these observations that the chromatin
of the small forms which have entered leucocytes divides with a
corresponding increase in the amount of cytoplasm.
Finally the larger forms burst, setting free small parasites
resembling closely the forms found in the red corpuscles. It seems
obvious, then, that the parasite undergoes a definite schizogony
within the leucocytes.
A gamogonous and an agamogonous generation, as described
by Gonder for Theileria parva , could not be followed out in the
parasite of the Echidna.
The occurrence of bacilliform parasites, cross forms and Koch’s
bodies places the parasite of the Echidna in the genus Theileria , and
the specific name of Theileria tachyglossi is proposed.
The Echidna from which the parasites were obtained was
infested with ticks— Aponomma dec or o sum (L. Koch)—and was
apparently healthy.
SUMMARY
A new species of the genus Theileria was found in an Echidna,
Tachyglossus aculeatus .
This parasite, for which the name Theileria tachyglossi is
proposed, resembles very closely Theileria parva. Koch's bodies
were found in great numbers in the internal organs, and to a certain
extent in the peripheral blood, and represent a stage in the life
history of Theileria tachyglossi in the warm-blooded animal.
REFERENCES
Gilruth, Sweet and Dodd (1911). Observation) on the occurrence in the blood of vaiious
animals (chiefly Monotremes and Marsupials) of bodies apparently identical with
Ana pi <um a marginal Theiler, 1910. Parasitology Vol. IV, pp. 1-6, 1 plate.
Gonder (1909-10). Report of Government Veterinary Bacteriologist. Transvaal 1909-10.
P . 69.
238
EXPLANATION OF PLATE XVIII
All the figures were drawn by means of an Abbe drawing
apparatus from preparations fixed dry and stained with Giemsa’s
solution. Zeiss apochr. homog. 2 mm., comp, ocular 18 except
figs. 15 and 16, in which comp, ocular 8 was used.
Figs. 1-3. Bacilliform parasites.
Fig. 4. Ovoid parasites.
Fig. 5. Comma-shaped parasite.
Fig. 6. Pyriform parasite.
Fig. 7. Round parasite.
Fig. 8. Cross-form parasite.
Fig. 9. Four parasites in one corpuscle.
Fig. 10. Anaplasma.
Fig. 11. Koch’s body, large free form with deep-staining cyto¬
plasm.
Fig. 12. Koch’s body with pale-staining cytoplasm.
Figs. 13, 14. Free parasites in leucocytes.
Figs. 15, 16. Intracellular Koch’s bodies.
THEILERIA TACHYG LOSSI.
239
AN INVESTIGATION INTO THE CAUSES
OF THE PREVALENCE OF ENTERIC
FEVER IN KINGSTON, JAMAICA ; WITH
SPECIAL REFERENCE TO THE QUESTION
OF UNRECOGNISED CARRIERS
BY
H. HAROLD SCOTT, M.D. (Lond.), D.P.H. (Hons.)
GOVERNMENT BACTERIOLOGIST ; PATHOLOGIST TO THE PUBLIC GENERAL HOSPITAL, KINGSTON, JAMAICA
{Received for publication I February , 1915)
Ten Charts
CONTENTS
I. Introductory .
II. Water Supplies .
III. Milk and other forms of Food .
IV. Flies.
V. Sewage Disposal .
VI. Carriers .
VII. Bacteriological Investigations :—
i. Method employed .
ii. Cases with post-mortem signs of enteric fever giving positive results
on cultivation of the bile.
iii. Cases giving similar results, but showing no evidence post-mortem of
enteric fever.
iv. Deductions from the fact of isolation of the Bacillus typhosus from
the gall-bladder or its contents .
v. Peculiarities of the organism isolated in some instances .
vi. Length of stay of the organism in the gall-bladder .
vii. Presence of bacilli with and without obvious inflammatory changes
in the gall-bladder.
viii. The question of atypical cases without characteristic changes
VIII. Personal Contact.
IX. Legislative Recommendations .
References.
Table .
Charts
page
240
2 45
2+7
248
249
251
255
256
158
261
262
265
265
266
267
268
27^
240
I. INTRODUCTORY
The^extensive prevalence of enteric fever in the town of Kingston,
Jamaica, and its suburbs has been for some years a cause
of considerable uneasiness, amounting at times to actual alarm.
This uneasiness is fully justified when it is considered that the city
is ideally situated in that it is exposed to bright sunshine practically
all day long and on nearly every day of the year, for the weather
might be regarded as that of perpetual spring and summer with
occasional showers. The disposition of the town is that of a gentle
slope from the upper parts right down to the sea, and therefore with
an ideal natural drainage. The obvious inference is that the reason
of the undue prevalence of enteric fever must be looked for in the
inhabitants themselves. Part of the town is provided with a water-
carriage system of sewage disposal, but a large part is not. This
part reminds one very much of the old time military camp, when
each fresh company of troops arriving, encamped upon or close by the
latrines of the last, or, when the troops were stationed in a place for
some time (comparable to the stationary population of Kingston),
they were from a sanitation point of view living on their own
dunghill.
Chart I, compiled from figures which have been given to
me by the kindness of the Acting Medical Officer of Health,
shows that the number of notifications of typhoid fever begins to
increase in March and reaches a maximum in June. A considerable
proportion of these, of course, are notified by medical practitioners
in the city without any bacteriological examinations, and con¬
sequently include many which are perhaps not enteric fever at all.
I think this inference is warranted, because if the figures are
compared with those from the laboratory, where specimens which are
sent up from suspected cases are examined (see Table I), it will be
seen that as a rule more than half those examined give negative
results. We may, therefore, take it that a certain fairly large
proportion of those notified as typhoid fever are not true cases of
the disease. I consider that I am further justified in saying that the
laboratory figures, although based on a smaller total, are more likely
to be accurate as regards the disease in question than those given by
the Medical Officer of Health as 4 notifications/ This must not be
2 4 I
implied as reflecting in any way on the diagnostic capabilities of the
medical practitioners here, but it is an undoubted fact that many
cases which exhibit a continued fever of obscure nature, though not
enteric, are diagnosed as such when no bacterial tests—Widal,
culture of the blood, faeces, urine, etc*,—are carried out. Hence,
although it is probable that few cases of true clinical enteric fever are
missed, the number notified would show an excess in the actual
prevalence of the disease in the clinical sense. This remark applies
generally to all towns. In hospital records, such as I have more
Chart I.—Notifications of .Enteric Fever cases in Kingston during 1914.
particularly to deal with, a different state of things may exist. Having
a pathological laboratory in the centre of the town, any doubtful case
is subjected to a blood examination, and thereby two errors may be
rectified.
Firstly, by this means patients with obscure febrile symptoms,
but not clinically indicative of enteric fever, might have to be
reported as such, owing to positive laboratory findings.
Secondly, suspected cases, which would be reported as enteric
fever because of this suspicion, are not so notified owing to negative
242
results of laboratory tests, or the notification is delayed till other
symptoms render the diagnosis clear.
Although these two points are antagonistic, the first tending to
increase while the second would lead to a diminution of cases
notified, the latter certainly preponderates, as is shown by the
appended Table I, where it will be seen that of the specimens sent
up some 50 per cent, proved to be negative. In other words, about
one-half of suspected cases are not suffering from enteric fever, that
is, the notifications, as stated above, are in excess of the actual
prevalence of the disease. Cases wrongly diagnosed as enteric
fever will, it is true, erroneously swell the notifications, but if labora¬
tory examinations of such are made free (see p. 269), these
diagnoses can be rectified later, and the Medical Officer of Health
can make the necessary alterations in his records.
One of the main dangers of a wrong diagnosis is that such cases
may then be traced to the operation of some factor in the spread of
typhoid infection which really has played no part, the case not being
one of typhoid fever at all. On the other hand, this is a very small
point in cpmparison with the overlooking of positive cases. In
other words, the erroneous diagnosis of a non-enteric case as enteric
is venial compared with the failure to diagnose a true case because
the clinical manifestations happen to be anomalous, through omitting
to make use of a laboratory when it is situated centrally, and thus
allowing a possible carrier to go about unwarned.
It must be remembered that among the 4 negative ’ cases are
included some which on subsequent testing yielded positive results,
since some specimens are sent up too early in the disease (second or
third day, perhaps), while in others agglutinin formation may be
delayed.
The taking of a sample of blood for a Widal test is simple and
causes practically no pain, but in the interpretation of the result
there are certain pitfalls. These have been dealt with in more detail
elsewhere (Scott, 1913), so it will suffice merely to mention the
following facts: —
(1) Though a positive result is obtained in the majority of
cases (95 per cent.) at some time or other of the disease,
there is, it would appear, a small percentage in whom it
is not given. Into the reasons put forward for this I need
243
Table I.—Showing the numbers of specimens sent to the laboratory month by month
for Widal’s test during the year 1914 , with the results.
Month Number Positive Percentage Negative Percentage Doubtful Percentage
sent
positive
1
negative
doubtful
January .
21
7
33
i ;
13
62
1
| 5
February ...
26
8
31
>7
«5
i
4
March .
54
2 3
42
2 9
54
, I -
2
4
April .
68
2 7
40
39
57
2
3
May .
"9
1
40
33
75
63
4
4
June.
109
59
54
47
43
3
3
J“>y .
62
22
35
39
6 3
|
1
2
August .
65
22
__
w
4 -
40
61
3
5
September ...
69
21
3°
46
66
3
4
" “ 1 _ 1 1
October .
7 1
19
2 7
5*
70
2
3
November ...
|
7*
24
34
43
61
4
5
December ...
73
21
2 9
49
67
3
4
244
not enter now—deficient agglutinin formation, excess of
agglutinoids, etc.
^2 That a positive result is in many cases, one might almost
say in most, not obtained until the end of the fitst week
of the illness, or even later.
(3) That in cases of exceptional severity the test may give a
negative result.
(4) That a positive result does not necessarily mean that the
present illness of the patient is enteric fever. Patients
who have been through an attack previously, it may be
years before, not uncommonly give agglutination,
especially if they are carriers, but by no means always so.
Blood culture, of course, is best for early diagnosis. The
percentage of cases from which the organism can be isolated
diminishes from 96 in the first week to 35 in the fourth; that is,
the value of the Widal test gains as that of the blood cultivation
diminishes.
To state briefly the effects of the Widal results on the notification
returns (in addition to what was stated before): negative results,
owing to early stage of disease, delayed reaction, and so forth,
would tend to diminish the number notified, while positive results
occurring in old, cured cases, suffering now from totally different
affection, would tend unduly to swell the returns.
Bearing these few facts and reservations in mind, the charts
giving the curves of notifications to the Medical Officer of Health
for this disease for the last seven years are most instructive (see
Charts II—X, pp. 279-284).
In nearly all of these charts it will be noticed that there is a
distinct rise in the number of cases notified in the second quarter of
the year, and in some years a small increase during November. In
1912 the November increase was markedly high (see Chart VI), the
reason for which I am unable to conjecture at this interval of time.
Comparing the average table of notifications (Chart X) or the 1914
one (Chart VIII) with the laboratory results for the same year
(Table I), the first-named increase is most distinct.
The epidemiological aspect of the question is a fascinating one,
but appertains rather to the domain of the Medical Officer of
Health, and I must pass on to the points which belong more to my
own investigations.
Chart X reveals the startling fact that for the past seven years
—I have not been able to obtain reliable statistics prior to that—
the average number of fresh notifications of enteric fever cases has
never fallen as low as 15 in any month of the year, while in May and
June the average for the same period reaches the high figure of 30
and over.
Briefly stated, then, the problem to be solved is two-fold,
namely: —
(1) Why is the permanent enteric fever prevalence so high in
a town favourably circumstanced as regards sunlight and
natural drainage?
(2) Why is the prevalence at its highest in the late spring and
early summer months of the year ?
I am strongly of opinion that the solution of the problem
of enteric fever prevalence, and thereby the hastening of the
desideratum of the final suppression of the disease, is more likely to
be reached by studies of the affection in areas and places where it is
endemic than by investigations of local acute outbreaks where of
necessity the investigation has to be, to a certain extent at least, a
hurried one. A town in which the average notification total in the
best months is between fifteen and twenty cannot be regarded
otherwise than as an endemic area.
In beginning an investigation into the causes of such prevalence
of enteric fever in Kingston, one’s thoughts were naturally first
directed to the usually recognised sources of the spread of the
disease, namely, food and water, flies, and dust.
II. WATER SUPPLIES
Bacterial examinations of the Kingston water supplies are
regularly carried out at this laboratory in order to safeguard their
purity. The supply is three-fold; of these, two are in constant use,
while the third is a subsidiary supply called into requisition in dry
months, when the others prove inadequate for the demands of the
population. If the analyses at any time give indications of
deterioration from the usual standard, which is a good one and well
246
up to that recognised for tropical waters, that is, if any indications
of excretal contamination are found, the fact is immediately reported
to the Central Board of Health and to the Kingston General
Commissioners who control the supplies, and steps are at once taken
to locate and remedy the mischief.
It came as a matter of great surprise, therefore, in June, 1914,
when the town was scared by a report from one in authority in
public health matters here, that the water supply, and especially the
auxiliary one installed comparatively recently at great expense and
after several careful analyses, consisted of ‘ diluted sewage/ and
could ‘by no methods be rendered fit for drinking/ and was the
cause of the large incidence of enteric fever.
Fortunately, to put the matter briefly, the report proved to be
quite unfounded, a mere canard, for the incriminated source of
supply was not, nor had it been for some time, in use, the other two
sources proving quite sufficient for the demands of the population.
Moreover, these latter showed no deviation from their customary
standards.
To discuss in detail the points brought forward would unduly
prolong this paper, so I will content myself with stating that against
the supposition of the disease in Kingston being water-borne are the
following facts:
1. The population of Kingston is estimated at 58,352, and the
notifications of cases of enteric fever include all those in the Public
General Hospital, which draws not only from Kingston, but also
from the neighbouring parish of St. Andrew, at all events the lowest
and most densely populated part of it. At the very smallest
computation, therefore, we may take it that the combined population
from which the notifications came was 60,000.
During May, the month in which the greatest number of cases
was notified, there were fifty notifications, that is, less than 1 per
1,000. This is a large proportion in a town with every natural
advantage for drainage, but a very small proportion as compared
with what would occur if the infection were water-borne, since over
58,000 individuals drink this water. The records of the Worthing
outbreak in 1893 give 557 cases in three weeks in a population of
15,000, or approximately 37 per 1,000. In the Maidstone epidemic
of 1897 there were 1,201 cases in three weeks out of a population of
247
33.830. or 35 cases per 1,000. In Lincoln in 1905 in the same period
approximately 11 per 1,000 of the population were attacked. In
Kingston, as already stated, it is less than 1 per 1,000.
2. The penitentiary, with a large number of inmates, and the
lunatic asylum of 1,400 patients, use the same water, and moreover,
water from no other source, yet amongst these during the same
period there were no notifications at all.
3. The different quarters of the town, though having the same
water supply, contributed in very different proportions to‘the total
of cases of enteric fever notified. Thus, in the month referred to,
from the N.E., N.W., S.W. and S.E. districts there were notified
respectively 11, 23, 7, and 5 cases, that is, 34 in the northern to 12
in the southern parts.
In subsequent months for which I have, by the kindness of
Dr. Cross well, the Acting Medical Officer of Health, been able to
obtain details, in July, out of a total of 19 for all four districts, 15
came from the northern and 4 from the southern; in August, 19 out
of 28 were notified from the same parts; in September, 12 out of 20;
in October, 11 out of a total of 17; in November, 12 out of 17. The
significance of these facts will appear later.
III. MILK AND OTHER FORMS OF FOOD
Personally, I have not observed any cases in Kingston in which
the infection could be definitely traced to contaminated milk supply.
This is a matter for the Medical Officer of Health, and is outside
my particular province. I am on sure ground, however, in stating
that here, as in most tropical countries, milk is very carelessly
handled, and the cleanliness of the vendors and their receptacles is
by no means above suspicion. Watering the milk is a common
practice, and if the tap is too far off, a nearer source is called into
requisition without compunction. One vendor has been seen to
abstract milk and fill up to the original bulk with liquid flowing
down the gutter at the side of the road. When it is remembered that
these same gutters constitute for the poorer inhabitants the State-
provided latrine, one source of infection is not far to seek.
Besides the faulty methods of handling and delivering milk,
another and more remote source of infection is possible. Few, if
248
any, of'the purveyors—I personally know of none, except possibly
at the Government farm—make even a pretence of sterilising milk
bottles or cans. So infection of milk may arise by washing the
various utensils with polluted water, by employment of carriers, or
of persons suffering from mild or ambulant forms of the disease.
As regards other articles of food, shell-fish is not eaten to any
extent in Kingston, and certainly not by the poorer classes.
Vegetables are brought into the town from neighbouring districts by
small cultivators, who deposit their excreta close to the huts in which
they live; they form a possible but not a proved source.
Sweets, cakes, and such like, are on sale at various dusty street
comers, and it is only recently that the vendors have been ordered
to keep these delicacies ( ?) under cover to protect them from dust,
flies, or the fingers of intending purchasers. These people commonly
take up one article after another before coming to a decision as to
which gives most value for their money. The hovels in which sweets
and foodstuffs are prepared are anything but above suspicion.
Indeed, I know of one case where the sweets thus sold were actually
being made in a hut in which an enteric patient was lying. This was
only discovered by the merest chance, and the tracing of infection to
its source when spread from unrecognised or unreported cases must
of necessity often be a matter of chance. Nevertheless, it is clear
that from the standpoint of preventive medicine, the unrecognised
cases are of greater importance than cases which are erroneously
reported as enteric fever.
IV. FLIES
These insects are very troublesome in some parts of the city,
particularly in the poorer parts whence many of the enteric cases are
notified. It is well known, in fact almost obvious, that the chances
of dissemination of infection by the agency of flies in a well-sewered
city are much less than in cases where proper care is not taken in the
disposal of excreta.
In Kingston the parts unconnected with the water-carriage
sewage system are those in which the majority of cases arise. In
these quarters flies are troublesome at most seasons of the year, but
the time when they become a positive pest is that of the 4 Mango
249
season/ starting about May, and this is the time at which the
enteric fever notifications begin especially to increase. This part of
the question is barely touched upon here; I hope to consider it
subsequently.
A fly census in different parts of the town, and the establishment
of a correlation between this and the districts whence notifications of
enteric fever are sent, would be an interesting matter, but is more
within the province of the Medical Officer of Health than that of
myself. With his consent, I would like to undertake such an
investigation at some future period, if my other duties will permit.
Dust, though often very troublesome in Kingston, is not regarded
as so potent a cause of the spread of enteric fever as has been
believed, especially in a tropical country with prolonged exposure
to the sun’s rays. In fact, it would be more correct to say that as a
direct cause dust accounts for very few cases, and that many of those
formerly attributed to dust, as air-borne, may be more reasonably
ascribed to the agency of flies.
House-flies have been captured in dwellings near badly-kept
privies used by enteric fever patients, and the bacilli have been
isolated from them. Experimental investigations (Hamilton, 1903)
have shown that living bacilli may remain in or on the bodies of flies
for twenty-three days after infection.
Cockroaches also, which, like the poor, are always with us in
Jamaica, swarm near badly protected food, and may easily act as
mechanical carriers of infection. Food protection, except from the
point of view of larceny, does not enter into the domestic arrange¬
ments of the poorer inhabitants of the city of Kingston.
V. SEWAGE DISPOSAL
A few words will suffice to sum up this section. The water-
carriage system is laid down for the lower part of the town only.
The upper parts, N.E. and N.W. districts, are largely furnished—
more or less badly—with privy middens, dry earth closets, and so
forth, nothing less than a standing and open invitation to flies,
which freely avail themselves of the opportunity.
Some of the huts inhabited by natives in Kingston have the
luxury of a privy to themselves. In other instances there is one to a
250
yard in which there may be four or more huts and as many families,
living as one small related community—very much related—so that
privacy is more honoured in the breach than in the observance. The
gutter passing the front doors is so handy that it is by no means an
uncommon thing to see children, and even adults, obey the calls of
nature in the open street. Again, natives come down from the inland
districts, walk 15 miles or more to bring produce to sell in the town,
stay the night in Kingston or on the borders of the city, and sleep
on the pavement at the side of the road. These people cannot be
blamed for depositing their excreta in the road or in the gutter, for
public latrines are not provided for them.
These itinerant vendors may account for a few cases, but if so the
association would be practically impossible to trace. I think that in
Kingston, considering the class of patients dealt with in hospital,
with whom my special investigations shortly to be related were made,
and the quarters of the town from which they came, there is but little
importation of the disease from outside. It must arise, therefore, to
a large extent, if not entirely, in factors whose operation is
localised.
It has been shown from observations (by Rosenau, Lumsden,
Kastle, and Anderson, 1909-11) extending over several years in the
district of Columbia, U.S.A., that there is in general ‘no striking
difference in the prevalence of typhoid fever in the sewered and
non-sewered districts/ but at certain seasons, particularly the time
when flies are numerous, the non-sewered districts showed a much
greater incidence.
Also, the chances of infection being conveyed to the healthy from
close association with the sick are found to be considerably greater
amongst those who dwell in residences provided with privies, than
amongst those whose residences have a water-carriage system
connected with the regular sewage system of the city.
In Kingston many of the privies are in a parlous state and far
below the modem standard of a sanitary privy, if such there be.
They are not water-tight, they are not protected from flies, even in
the better houses they are often situated close to the kitchens and
even dining and sleeping rooms. The kitchens are not screened,
and flies can travel readily from the privy to the food. Also, as
already stated, in the poorer quarters one privy may have to serve
for several families, so that many individuals may become infected
from a single carrier or a common source.
There is always a danger, in investigating for one single cause,
of ignoring or forgetting other contributing causes. Thus, an investi¬
gation at one time might elicit evidence pointing strongly to one
factor, e.g., flies, while at another the evidence might be to the
incrimination of some article of diet—milk, shell-fish, etc.—and tend
to minimise the importance of the former. But a high general
typhoid rate with increased prevalence at some particular season of
the year makes one think firstly what general and continuously-
acting factor is at work; and secondly, what factor or factors are
more strongly operative at the time of greater prevalence, and
whether the reason is to be discovered in an increased scattering of
seed, increased receptivity of soil, or both combined.
In Kingston, where the variations of temperature are compara¬
tively small in the different seasons and where perpetual summer
reigns, the following factors must be considered: prevalence of
flies at certain times, diminution of rainfall and concentration of
pollution, or sudden heavy rains washing pollution from the banks of
a stream into the supply, the ingestion of raw fruits, and the
prevalence of other intestinal affections rendering the subjects more
liable to contract typhoid fever when exposed to infection, and so
forth.
The water supply, the fly nuisance, the sewage arrangements,
are matters for the Medical Officer of Health to consider and deal
with. I could not interfere with them without his full consent and
co-operation. But even granting that these were unsatisfactory,
there remains still unaccounted for the source whence the flies, or the
food, or the dust obtained the organisms of the disease, and carriers
are naturally suspected.
VI. CARRIERS
The question of carriers is a matter with which my work as
Government Bacteriologist is closely connected, but there were
certain difficulties in the way of investigation to avoid interference
with the work of the Medical Officer of Health.
There are difficulties also as to obtaining material, for persons in
health cannot be prevailed upon to send excreta in order that it may
be determined whether or no they are typhoid carriers. The
objection is quite a natural one, and more particularly would this
be the case when the finding would lead to certain rules and restric¬
tions being imposed. Moreover, it is fairly certain that, whatever
rules were given and however mild the restrictions, they would be
evaded in some way or other. A rule without an enforceable penalty
for disobedience is better left unmade. The native who would be
found to be so altruistic as voluntarily to place restrictions upon
himself for the good of others would indeed be a ‘rara avis.’
Further, if the results of examination were negative, the case would
by no means be proven, as the carrying might be intermittent.
The question of the percentage of enteric fever patients who
become carriers has been studied by many investigators. The
results, so far as the literature at my disposal goes, may be briefly
given. Lentz (1905) states that about 4 per cent, of typhoid patients
discharge the bacteria in their excreta at least ten weeks after
convalescence. Schneider (quoted by Besanqon) estimates the
chronic carriers as 3 per cent, of patients. Lehmann and Neumann
give 2 per cent, to 4 per cent. Nieter and Liefmann (1906-7) isolated
7 out of 250 patients (that is, 3 per cent.) in an asylum. Asylums
generally appear to harbour an abnormally high proportion of
carriers.
But cases excreting only for so short a time as ten weeks after
the illness cannot be regarded as carriers in the real sense of the
term. The percentage of chronic carriers is variously estimated at
from i‘5 per cent, to 5 2 per cent. So far as I have been able to
ascertain, the following figures are authentic: —
Out of 400 cases of enteric fever 6 excreted the bacilli for more
than three months, giving a percentage of 15; of 316 cases 9 carriers,
or 2 8 per cent., were found; of nearly 7,000 cases 166, or 24 per
cent; while of the large number of 11,000 cases 411 carriers were
found, a percentage of 37.
Altogether I have found results recorded of 18,431 patients, and
of this large number 59 2 were detected as carriers, a proportion of
3 21 per cent. We shall be safe in taking it that, as an average,
some 3 per cent, of patients become carriers for three months at
least.
253
Of i,800 healthy subjects reported upon by Kayser in 1907 giving
no history of previous typhoid fever, 27, or 15 per cent., were found
to be excreting the bacilli. In Columbia the excreta from 1,000
healthy persons were examined, and of these three were found to
contain the organisms; one of these excreted them in the urine, the
other two in the faeces; all three were males.
Returning to the condition of things in Kingston, my suspicions
were that, as the enteric rate is so high, there might be individuals
going about apparently in good health, who were unwittingly
spreading the infection, or that the percentage of carriers here is
greater than has been found to prevail in most places.
VII. BACTERIOLOGICAL INVESTIGATIONS
It is now an established fact that during an attack of enteric
fever the bile frequently becomes infected; this is corroborated by my
results as shown in the table of cases so examined (see Table II).
If now, as is also known to occur, an inflammatory process arises
in the gall-bladder, the bacilli may be discharged either continuously
or at intervals from the bladder into the intestines for periods of
almost indefinite duration.
Progressing a step further, it may be said that in many cases
the chronic bacillus-carriers are individuals harbouring the typhoid
bacilli in their gall-bladders.
I determined, therefore, to make cultivations from the bile of
every patient dying in the general hospital—regardless of the actual
cause of death as diagnosed clinically or discovered post-mortem—
in order to find out whether any, and if so how many, were carrying
the organisms in their gall-bladders at the time of death.
Of course, in a large percentage of patients admitted with a
history of enteric fever and dying from this disease, and also in
patients wrongly diagnosed, but actually suffering from enteric fever,
one would naturally expect to find the organism. But there was
yet another class for which I particularly wished to look out, namely,
patients who were admitted into hospital for some affection totally
unconnected, or not suspected of being connected with enteric fever,
and who gave no history of any previous attack of the disease or of
any prolonged fever which might be so explained. These persons,
* 5 +
by harbouring the Bacillus typhosus in their gall-bladders, had
possibly, if not probably, been acting as carriers, and, had they
chanced to recover, would have again gone about spreading the
organisms broadcast. These were the individuals who constituted
a danger to the community, and if such were found to exist, the fact
would go far to account for the undue prevalence of enteric fever in
Kingston.
It is but right to call attention to the fact that the mere denial
on the part of the patient as to his having suffered previously from
the disease does not exclude the possibility of his having passed
through an attack. The native is so accustomed to short attacks of
malarial fever that any ‘ fever * of a prolonged or severe character
usually makes a permanent impression on his mind. When he speaks
of a 1 bad fever * lasting for two or three weeks, this is quite sufficient
to enable it to be stated that the history evinces a strong probability
of the patient having had an attack of enteric fever in the past.
The investigation into the results of bile cultivations is still being
continued. Up to the present 120 cases have been dealt with,
sufficient for a preliminary note, and sufficient to form a basis for
discussion (see also Addendum, p. 269). Before entering more
fully into a consideration of the various points which present
themselves, I would beg to call attention to the appended list of
cases where the results are briefly recorded (Table II, pp. 272-278).
The following points worthy of more detailed mention present
themselves on a persual of this table:
1. The number of cases exhibiting post-mortem the signs of
enteric fever from which a positive result was obtained
from cultivation of the bile, and arising out of this, the
number of cases with similar signs from which the result
was negative.
2. A corollary to the last, namely, a consideration of the
cases which showed no signs of enteric fever, but from
which a positive culture was obtained.
Other points of interest which declared themselves during the
investigation were: —
1. Peculiarities of the isolated organism in certain instances.
2. Absence in most cases of macroscopic signs of inflamma¬
tion of the gall-bladder.
255
(i) The method of procedure employed in each case
As soon after death as possible, before starting on the usual
post-mortem routine, the gall-bladder was exposed, the surface was
sterilised by searing, and the organ held by sterile forceps. A small
incision was made with sterilised scissors. Tubes of peptone broth
were ready containing quantities of a i in 10,000 solution of brilliant
green, varying from 003 c.c. to 3 c.c., namely, 0*03, 006, 012,
0*18, 2, and 3 c.c., the brilliant green being added immediately
before the tubes were taken to the post-mortem room. A loopful
(4 mm. in diameter) of the bile was introduced into each tube. Six
tubes were sometimes used, but on no occasion less than three,
because it was early noticed that the brilliant green had a variable
inhibiting power, not always proportionate to the amount added.
In other words, some strains of the bacillus obtained showed less
resistance to the brilliant green than did others.
The tubes after inoculation were placed in the incubator at 37°C.
and examined on the following day. If there was but a slight
growth with faint cloudiness of the medium, further examination
was postponed for another twenty-four hours. If there was no
growth by that time the tubes were kept for a week before the bile
was noted down as being ‘ sterile.*
If, and when, growth occurred a loopful was plated on to
Rebipelagar for isolation, and suspicious colonies, non-lactose
fermenters, were transferred to nutrient broth. Secondary tests were
then carried out with various sugars, namely, glucose, saccharose,
lactose, maltose, mannite, dulcite, and sorbite.
Any organism isolated, which gave the usual typhosus reactions,
was then tested for its agglutination power with an immune serum
which I prepared, and which gave an agglutination titre of 1 in
4,000 with its own producing strain. Only those organisms which
gave a positive result with the serum diluted to about this degree
were regarded as true Bacillus typhosus. As is well known, to test
with the serum of a typhoid patient or convalescent is quite
inadmissible for the identification of a suspected bacillus. In such
a serum besides the main specific agglutinins for the Bacillus
typhosus , there may be present a whole series of group-agglutinins,
agglutinoids and iso-agglutinins, in such concentration a$ to con¬
stitute a source of gross errors.
2 5 6
In the investigation, control tests were carried out with
corresponding dilutions of normal serum of the derivative animal,
to exclude the chance that the agglutination observed with the
immune serum might be an action of a normal serum on the strain of
bacterium under examination.
A second control was also put up, using normal saline in place
of the serum,' in order to exclude any incidental agglutinating
action of the diluent—a pseudo-agglutination—which might appear
from spontaneous clumping or insufficient emulsification.
Bruns and Kayser (1903) propose to use only a low dilution,
such as 1 in 100 of the immune serum, but this course is liable to
lead to erroneous conclusions, which need not be fully discussed
here. A long series of dilutions has been made in each of my cases
in order to determine in what degree of dilution of the serum the
agglutination would take place. As Pfeiffer, Lipschiitz (1904),
and others have pointed out, only when this corresponds fairly
approximately with the estimated titre of the serum ought the
a ggl u ti na tio n to be regarded as evincing the true typhosus nature
of the organism in question.
(ii) Cases with post-mortem signs of enteric fever from which a
positive result was obtained on cultivation of the bile.
Out of the one hundred and twenty cases, twenty-eight were
shown at the autopsy to be suffering from enteric fever, that is to say,
definite macroscopic lesions of the disease were present. In twenty-
four of these the Bacillus typhosus was isolated from the bile, and
in one other, whose blood during life had given a positive agglutina¬
tion with B. paratyphosus A, and negative with B. typhosus , and
who presented symptoms typical of enteric fever, the B. para¬
typhosus A was isolated. We may therefore say that from twenty-
eight cases of enteric fever the causative organism was isolated from
the bile in twenty-five, or in 89 28 per cent., although I am aware
that the reckoning as a percentage with so few cases as twenty-eight
is liable to error. This percentage number agrees almost exactly
with the figures of Forster and Kayser (quoted by Hewlett), who
obtained pure cultures from the gall-bladders of seven out of eight
cases. A better comparison may be made between these cases and
257
mine if we divide my twenty-eight into series of eight. For the
first eight I obtained the B. typhosus in pure culture six times, and
twice the organism was associated with a lactose-fermenting one,
which proved to be the ordinary Bacillus coli. The B. typhosus
was thus isolated in all the first eight. One of the cases in which
the B. coli was present also died from general peritonitis following
perforation of a typhoid ulcer, and the bile may have become
infected by general systemic infection, or, what is an equally
probable explanation, the B. coli may have entered as a contamina¬
tion in making the culture. If the former explanation is the correct
one, the fact shows that the bile and brilliant green do not always
have an inhibiting action on Coli organisms.
With regard to the second series of eight, I obtained the
B. typhosus in pure culture in six, and B. paratyphosus A in
another, also in pure culture; in other words, the organisms were
present in pure culture in seven out of eight. In the third series,
B. typhosus was obtained in pure culture in six, one remained
sterile, while from the other B. coli only was obtained.
The remaining four of the twenty-eight yielded the B. typhosus
alone in each case.
Hiss and Zinsser (1910) in their text-book merely state that the
‘ typhoid bacilli have been frequently observed in the gall-bladder at
the autopsy. 1 They do not mention how frequently. According to
Meyerstein (1907) the bile of typhoid patients nearly always
contains the Bacillus typhosus , though its presence does not often
give rise to clinical symptoms. Its presence may, however, set up
inflammatory processes in the wall of the gall-bladder. Thus
Stewart (1901) found a condition of cholecystitis in about 1 per
cent, of cases. Similar findings have been reported by Forster and
Kayser (1905), Hilgermann (1909), Findlay and Buchanan (1906),
Marmann (1908), Nieter (1906-7), and others.
The presence of the bacilli in the gall-bladder of patients
suffering from enteric fever is not to be wondered at when we
consider that the disease is, in the early stages at least, a
bacteriaemia; that experiment has shown that it is by way of the
blood-stream that the gall-bladder becomes infected, and that the
conditions prevailing there appear to have a favourable influence
upon the growth of this organism. For it has been shown by
Blackstein and Welch (1891) that after intravenous injection of the
bacilli into guinea-pigs they were recognisable in the bile four
months later, when all other organs appeared to be completely free
from them, while after subcutaneous, intraperitoneal, and intra-
gastric inoculation the bile remained sterile, though it was infected
after intravenous injection, even with the cystic duct ligatured
(Doerr, 1905). This appears to be the usual mode of infection of
the bile in man, namely, by way of the circulation. It is only fair,
however, to mention that Koch and Chiarolanza (1909) have held
that there is an alternative route direct from the intestine to the bile
through the ductus choledochus, at all events in experimental
infection of rabbits. In the case of the human subject, however,
this latter is very unlikely, seeing that the lower part of the small
intestine is that most affected, and it would indeed, with respect to
the local condition, be a 4 long way to go* when the route via the
blood-stream is so much more accessible and rapid. Moreover, the
bile appears to be infected so very early in the disease.
(iii) Cases which showed no post-mortem evidence of enteric fever ,
and in which no history of such was obtained , but which t
nevertheless , yielded a positive result on cultivation of the
bile.
This group is of the greatest importance in lending support to
the suspicion on which the investigation was undertaken, namely,
that unrecognised possible carriers are going about in Kingston in
larger proportions than have been estimated in other countries.
As has been already stated, some 3 per cent, of patients
become carriers for a considerable time. Of this first series of
one hundred and twenty autopsies there have been found four who
up to the time of onset of their final illness had been going about
apparently in perfect health, who gave no history of having had an
attack of typhoid fever. In one case, No. 2, there was an indefinite
history of prolonged fever which might possibly have been typhoid,
though from the symptoms and post-mortem signs this might equally
well be ascribed to tuberculosis. The four mixed freely with their
fellows, and lived in the poorer, insanitary, and unsewered parts
of the city.
*59
These cases are worth quoting briefly:
1. C. C., male, aged 49 years, white, No. 7 in the appended
list, Table II. Admitted to hospital 4 p.m. March 31, 1914*
a history of having at 9 a.m. ‘taken rat-poison in mistake for
phenacetin.' He died atr 11.40 p.m. The fact that more than 400
grains of arsenious acid were found in the stomach and viscera, even
after the vomiting which had continued for ten hours, points to
deliberate suicide, for no one would take nearly an ounce ol
phenacetin for a single dose. His neighbours stated that he went
in and out amongst them apparently in perfect health till the day
of his death. In bile culture B. typhosus was obtained.
2. B., a Coolie woman, aged 21 years, No. 34 in the list, when
admitted complaining of ‘fever, cough, and pain in the right side
of the chest.' Expiration was prolonged, and there were rhonchi
audible over both lungs.
The diagnosis of phthisis was made, and the patient died early
the following morning.
At the autopsy the lungs showed minute scattered nodules with
caseous contents, possibly tuberculous, but the chief finding of
interest was that the gall-bladder had practically disappeared, and
there was an ulceration into the duodenum just beyond the pylorus,
with a gall-stone about the size of a small hazel nut in the aperture.
There were three abscesses in the liver, one cavity being the size
of a tangerine orange.
From the nucleus of the gall-stone, the only one found, a pure
culture of B. typhosus was obtained.
3. S. D., male, aged 19 years, black, No. 73 in the list, was
admitted with a history of six days* ‘ cough, pain in the right side
of the chest, and fever.' The percussion note over the right lung,
lower lobe, was quite dull. ‘ No breath sounds could be heard, but
vocal fremitus and resonance increased.' Widal reaction was
negative. The patient died ten days later.
At the post-mortem the right lung was quite solid; the upper
and middle lobes in a state of grey hepatisation, the lower lobe in
a condition of purulent infiltration, and in one place broken down
to abscess formation. The lung was large and heavy, and the liver
so displaced that the upper margin only reached one and a half
fingers'-breadth above the costal edge.
2 6 o
This, then, was a case of unresolved lobar pneumonia. No
history whatever was obtained pointing to enteric fever, but a culture
of B. typhosus was yielded by the bile.
4. E. R., female, aged 21 years, black, No. 113 in list, was
admitted to hospital on October 26th, 1914, with dyspnoea fend
general oedema, especially of the lower limbs. The urine was
albuminous and contained granular and hyaline casts. Death took
place on the following day.
The autopsy revealed a condition of chronic nephritis, the renal
capsules were adherent, the surface of the organs granular, the
cortex narrowed to about half the normal ratio, and there were two
small cysts in it. The heart was enlarged, the left ventricle being
hypertrophied, and the weight of this organ when empty was 480
grams. There was no sign of enteric fever found, and no
history obtained of any such disease. A pure culture of B. typhosus
was isolated from the bile, which agglutinated ‘after three sub¬
cultures with the typhoid immune serum in as high a dilution as
1 in 5,000.
Exclusive, therefore, of cases showing evidence of enteric fever
at the autopsy, the bacillus has been isolated from the bile of four
subjects:
1. Dying from arsenic poisoning, suicidal.
2. Dying from multiple abscesses of the liver.
3. Dying from pneumonia.
4. Dying from chronic nephritis and heart disease.
It would hardly be fair to draw conclusions from so few cases as
one hundred and twenty, and the investigation is being continued
(see Addendum, p. 269). All that can be said at present is that,
apart from patients treated for enteric fever at the hospital, and
apart from cases showing signs of this disease post-mortem, there
have been three among the first one hundred and twenty who were
harbouring the Bacillus typhosus in their gall-bladders at the time
of death. In a fourth who had done so, the organism was found in
a gall-stone which had ulcerated into the duodenum.
If we deduct the number of those who were suffering from the
disease, showing definite signs of it at the autopsy, we may state
that out of ninety-two subjects there were four who were harbouring
the organism of enteric fever.
26i
(iv) The deductions which may be drawn from the tact of isolation
of the bacillus from the gall-bladder or its contents
1. The subjects are chronic typhoid carriers. This is possible,
but could not be regarded as proved unless repeated examinations
of the excreta yielded positive results. In my cases this was
impossible, as they did not come under my observation till after
death.
2. The subjects are temporary carriers.
(i) They may, in the absence of history, have passed through
an attack of enteric fever and forgotten or not have known the fact,
either because of the mildness of the infection, or because of time
elapsing, or because the illness had been wrongly diagnosed. The
following cases show that the last may certainly occur:
No. 4, diagnosed lobar pneumonia.
No. 5, diagnosed pneumonia.
No. 9, diagnosed pneumonia.
In these three, typhoid ulceration was found but no signs
of pneumonia.
No. io, diagnosed general peritonitis. This was true, but
the peritonitis arose from the perforation of an enteric ulcer
which had not been suspected.
No. 88, diagnosed cerebral haemorrhage. There were the
usual signs of enteric fever, but none whatever of cerebral
haemorrhage.
No. 99, diagnosed pneumonia. This was present, but there
had been no suspicion of enteric fever, which was evident with
its typical pathological changes.
(ii) They may be ‘ porteurs sains * in the sense of never having
had the disease. In this connection I may quote an interesting case
which came to my notice as Government Bacteriologist more than
a year ago. It was mentioned in my paper already referred to, and
published in the ‘Practitioner* for November, 1913:
‘ A child of nine months suffered from an attack of typhoid fever, proved by
isolation of the bacillus. The question then arose as to the method by which
the child could have become infected. The little patient’s mother was the nurse
in charge of the enteric ward at a general hospital in the island, and it was thought
that she might possibly have taken some milk or other food from the ward home
to the child. A specimen of the mother’s blood was asked for. and it was found
to give a very marked agglutination of bacillus typhosus in high dilution.
262
‘ The nurse was perfectly certain that she had never suffered from typhoid
fever, or, in fact, from a prolonged fever of any kind ; she had always been healthy,
and “ never remembered being ill in her life ” (to quote her own words). The
next step was the examination of her stools and urine, and the bacillus typhosus
was isolated from the latter ; in short, this nurse was definitely a carrier, though
never having herself suffered from the disease, and she seems without doubt to
have conveyed the disease to her child.’
If the subjects are ‘porteurs sains * in the sense mentioned above,
that is, the organisms are present without setting up the disease, we
may find here support for the X Y Z theory of Pettenkofer. Thus,
it is possible that the bacilli may be harboured in certain situations
(in my cases the gall-bladder) for a lengthy period, but until the
other components of the aetiological complex are superadded no
disease results. We may, perhaps, compare it to the lowering of
resistance to the Bacillus tuberculosis , allowing development of the
organism and production of the disease.
Against this in my cases is the fact that two, possibly three, of
the subjects were considerably debilitated by illness, and there were,
nevertheless, no signs of enteric fever supervening. This argument,
it must be confessed, is of the nature of a two-edged sword, for,
firstly, mere debility by any illness may not be the Y or Z of the
aetiological complex, but some element more specific in nature may
be necessary. Secondly, some of the cases which showed signs of
enteric fever post-mortem, although a different diagnosis had been
made clinically, might be instances of the supervention of enteric
on a previous debilitating disease. This, again, opens up the
interesting question as to whether cases of enteric fever arising in
patients who have been in hospital for some time, a month perhaps,
may not in some instances be of f spontaneous * or ‘ autogenous *
origin, instead of being regarded as cases of ‘ contraction of disease
in hospital/ By the terms spontaneous or autogenous I imply the
‘ porteurs sains * who develop the disease after some other debilitating
illness, for example, typhoid fever following dysentery, and not
merely ordinary cases with a prolonged incubation period.
(v) Peculiarities of the organism isolated in some instances .
1. Motility. As has been noted by Fischer (1909), I have found
several times that when freshly isolated from the body, that is, from
the bile in my cases, the bacilli were but slightly motile, and in some
263
instances were devoid of motility altogether. But he goes on to
say that even after prolonged subculture the motility was only
slight. This has not been my experience. After the third or fourth
transference to nutrient broth the normal motility was always
regained; or by transference from agar to broth alternately.
I have not so far had to employ any of the special media for this
purpose, such as Losener’s serum, Terni’s peptone-free glycerin
bouillon, or the 2 per cent, glucose broth of Germano and Manrea.
2. Agglutinability. In my cases I have not yet met with an
organism which, while giving the cultural tests and sugar reactions
of the B. typhosus , was completely non-agglutinable with the
prepared immune serum. In several instances, however, the first
subculture showed very slight agglutinability, but, after sub¬
culture to the third or fourth generation and subsequently, the
organisms proved normally agglutinable.
In two cases I found a mixture of two typhosus colonies, one
easily agglutinable, the other only agglutinating with difficulty,
occurring from the same subject. A further point was also
observable, namely, that the former—the one readily agglutinable—
showed marked motility, while the latter—the poorly agglutinating
strain—was practically non-motile. Sub-cultivation, however, on
ordinary agar and in broth alternately, produced organisms equally
motile and equally agglutinable in the two cases. Thus, it is seen
that the diminution or absence of motility and a low degree of
agglutinability of a newly isolated strain of the organisms go hand
in hand. By further observations it was found in carrying out the
agglutination reaction that time was an important factor in some
cases. Thus, a strain which proved to be agglutinated to a slight
degree only within the ordinary time limit of two hours, would in
some cases, not in all, be strongly agglutinated if left for several
hours—up to 24—at room temperature, less if kept in the incubator
at 37 0 C.
Two of the explanations, which have been put forward to account
for the low degree of agglutinability of typhoid bacilli freshly
isolated from the body, are worthy of being mentioned in greater
detail; namely, that of Bail (1901), Muller (1903), Kirstein (1904),
and others, and that of Nicolle and Trenel (1902).
Their findings, some of which I have been able to confirm, may
be briefly stated thus : —
264
A culture of typhoid bacilli agglutinable in high dilution of the
immune serum was inoculated intraperitoneally into a guinea-pig.
After remaining a few hours the agglutination test was again
applied, and the titre of agglutination was reduced from the previous
1 in 4,000 almost to nil; there was incomplete clumping even with
1 in 300. This has been explained by supposing that the bacteria
which have been inoculated seize upon the agglutinophores which
possess the haptophore groups but not the zymophore, and thus put
them out of action. Bail (1901) similarly explains the reduction of
agglutinability of bacilli growing in a diluted immune serum, for the
normal degree of agglutinability is restored after a few transferences
back to ordinary nutrient media. It is but right to state, however,
that Muller (1903), Hamburger, and others regard the latter
phenomenon rather as a definite attempt at immunisation of the
bacillus against the injurious action of the immune serum, though
they are all agreed as to the subsequent restoration to normal agglu¬
tinability. It must be noted that this reduction is not marked in all
strains. We may thus briefly state this view: Under certain
conditions typhoid bacilli present in a patient seize upon the
agglutinoids and so lead to the production of cultures of the
organism which are but poorly agglutinable.
The second explanation rests upon the findings of Nicolle and
Trenel (1902) that cultures of the Bacillus typhosus , normally motile
and readily agglutinable, lost these properties when grown at 42 0 C.,
though they regained them on subsequent cultivation at 36° C.
They inferred that the high temperatures which some typhoid
patients exhibit might produce a similar change in the bacilli,
reducing their agglutinability. Higher temperatures still, such as
50°C., on the other hand, appear to bring about a hastening of the
agglutination and to render it more complete.
Eisenberg and Volk (1902) have shown by their experiments that
the non-agglutinable or poorly agglutinable strains may be,
nevertheless, identified by means of the immune serum. Heating the
cultures of typhoid bacilli or treating them with weak acids deprived
them of their agglutinability. Bacilli so treated are capable of
binding large amounts of agglutinin, hence the inference that such
treatment renders nugatory that part of the agglutinable substance
which is actively concerned in agglutination. The haptophore
group, however, is still present, for it is found that if such altered
z6s
bacilli be brought into contact with a typhoid immune serum all the
typhoid agglutinin can be removed, so that it will no longer
agglutinate any typhoid bacilli. This, as Wassermann has
suggested, might be usefully employed for the identification of
typhoid bacilli in those cases in which there is a suspicion that the
organisms have, owing to extraneous influences, lost their normal
agglutinability. But, seeing that cultivation again on ordinary
nutrient media soon restores the normal agglutinability, this method
of Wassermann would, or need only, find application where a
speedy diagnosis is urgently called for.
(vi) Length of stay of the organism in the gall-bladder.
As regards the length of time during which the bacillus can exist in
the gall-bladder, the limit has not yet been fixed. Cases of 16, 17, and
20 years have been reported by Zinsser (1908), Droba (1899), and
Hunner-Writer (1905), respectively. I myself related the case of a
faecal carrier of Bacillus paratyphosus A in 1911, whose primary
attack occurred in 1889, twenty-two years previously. Cases
reported by Dupre, Ramond, and Faitout, are quoted by Besan^on,
but it must be remembered that all these refer to the duration after
an attack of the disease. The four mentioned in my table, Nos. 7,
34, 73» and 113, gave no history of any previous attack at all, and
the time in them is impossible to determine.
(vii) Presence of the bacillus with and without obvious inflam¬
matory changes in the gall-bladder.
Doerr (1905) has stated that the bacilli make a prolonged stay
in the gall-bladder only if an inflammatory condition of the mucous
membrane is set up. In two of my four cases I could not make out
any such condition. It is possible, therefore, that their histories
were correct, that they had not suffered from the disease and that
the gall-bladder was merely their temporary sojourning place, a
port of call, as it were, in a ‘porteur sain,* as the bacilli were on
their way to being excreted. It may be incidentally noted that
gall-stones are very rarely found post-mortem here. Of all the
autopsies carried out by me during the last four years, I have only
found them to be present in two instances.
266
(viii) The question of atypical cases without characteristic changes.
Some of those from whom the organism was isolated, although
no symptoms were present and in whom there were no post-mortem
sign of the disease, might, of course, be examples of the atypical
cases without characteristic changes. Under this heading would
come such as have been reported as cases of 1 typhoid septicaemia *
If by this term we understand 1 a systemic affection in which the
causative organism not only gains entrance to the blood-stream but
also multiplies therein/ I know of no recorded positive findings of
such a thing in enteric fever, but an ordinary bacteriaemia is the
normal condition at an early stage of the disease. Absence of
evidence of intestinal affection, in spite of finding the bacilli in the
blood, is less uncommon. Such may be explained by the fact that
all transitions occur between the marked pathological changes in the
intestines and a minimal, scarcely recognisable lesion, and
this also apart altogether from the severity of the case. Also
de Grandmaison (1900) and Krokiewicz (1908) have produced
evidence that in several such cases definite formation of specific anti¬
bodies may be recognised in the serum. A second explanation is
that mentioned in the last section, namely, that the bacilli have
gained entrance into the body of a patient suffering from totally
different disease, the subject acting the part of a passive carrier or
receiver, as, for example, the entrance into the blood-stream of
typhoid bacilli accidentally by way of a tuberculous enteritis, as in
a case recorded by Busse (1908).
We must never lose sight of the fact that enteric fever is not an
intestinal infection. It is, on the contrary, a general one with
localisations of the bacteria in various situations, most frequently in
the intestine, but often in the liver and biliary system, sometimes in
the lungs, the bladder, the pleura, the meninges, and so on. Thus,
there may be a considerable variety ranging between a general
bacteriaemia without local infection specially marked in the
intestines, and more definitely localised forms with symptoms of
meningitis, broncho-pneumonia, cholecystitis, nephritis, arthritis,
etc. Yet again the typhoid nature of the condition may be
masked by accompanying infection, such as a streptococcal, or
tuberculous one.
VIII. PERSONAL CONTACT
With the histories often faulty, sometimes unintentionally, some¬
times deliberately so, it is an almost impossible matter to trace the
extent of personal contact and gauge how far it plays a part in
spreading the disease in Kingston. The poorer inhabitants live
often closely massed together, and many cases may thus arise from
a common source of infection, as has been already stated.
When the large number of possible sources of infection furnished
by typhoid carriers and by typhoid patients are considered, and
the many ways in which infection may be conveyed from the diseased
to the healthy, it can only be wondered why there is not more
typhoid fever rather than why there is so much.
If the extensive prevalence of this disease is to be coped with in
anything like an adequate manner, typhoid convalescents must be
subjected to greater restrictions than they are at present.
Particularly is this necessary with cases who pass cloudy urine
(bacilluria). They must be restricted until their excreta have been
examined bacteriologically and found to be negative. They must
not be freed entirely from observation even then, because of the
possible intermittency of the condition. As everyone knows, it is
the usual custom to discontinue all supervision over enteric fever
patients as soon as they are able to leave their beds. Consequently
in the majority of cases (I know there are exceptions, for instance
in the army, but otherwise the exceptions are rare) disinfection of
stools and urine is stopped as soon as the convalescent is able to be
up and about. To recognise how very unsafe this procedure is,
we have only to call to mind the results of examination of the excreta
of convalescents, and note the length of time during which many of
them continue to harbour the organism in question.
Though it is of great importance and value to discover the main,
or even one of the main, factors causing the spread of enteric fever
in a district, particularly in a crowded town like Kingston,
nevertheless, since the typhoid rate of incidence may be looked upon
as a good sanitary index of the hygienic condition of a community,
coincident improvements should be carried out in all the branches of
sanitation, if the prevalence is to be materially reduced. The
purity of the water supplies must be safeguarded, the sewerage
268
system should be modernised and the food supplies protected. The
patients should be kept under observation for a more prolonged
period than under present circumstances here (and so far as I am
aware the same remark applies elsewhere). More care should be
exercised both as to site for and manner of disposal of refuse.
Measures should be taken to minimise the prevalence of flies. The
community should be educated to greater personal cleanliness. In
short, all the conditions of general sanitation should receive attention
where such are found to be defective.
IX. LEGISLATIVE RECOMMENDATIONS
Having pointed out some of the various ways in which the undue
prevalence of enteric fever may be accounted for in Kingston, it only
remains for one to suggest means by which the possibility of spread
may be to a large extent controlled. The following brief recommen¬
dations do not include the measures which have been already
indicated, but apply rather to the legislative aspect of the question.
1. Rules should be made for early notification of all suspicious
cases. Much valuable time is lost if the physician waits
until he is practically certain, often a matter of a week and
sometimes of a fortnight or more. The form of notification
should be different from the usual one of 1 In my opinion
A.B. is suffering from enteric fever/ and might read thus:
‘ In my opinion A.B. shows evidence of symptoms which
may indicate enteric fever in an early stage/ Seeing that
the diagnostic ability of physicians is so variable, and
that the symptom-complex regarded as pointing to enteric
fever is so differently interpreted by individual medical
men, the regulation might be made that any patient with
unexplained fever of three days’ duration should be so
reported.
2. Blood cultures, if the case is in an early stage, or Widal’s
reaction after, say, the end of the first week should be
carried out at the Pathological Laboratory free of charge
to the patient.
3. Isolation of the patient should be required. In circum¬
stances where this cannot be carried out at home to the
269
satisfaction of the health officer, removal to hospital with
this end in view should be made compulsory.
4. If the patient is allowed to remain at his home, the health
officer must be satisfied that adequate disinfection of the
excreta not only can be, but is carried out.
5. No person continuing to live in the same dwelling as an
enteric patient should be allowed to prepare, or manufac¬
ture foodstuffs (including beverages) without the permission
of the health officer, given in writing.
6. Any room which has been occupied by an enteric fever
patient must be properly disinfected before being again
occupied.
7. Patients should remain under observation of the health
officer until the excreta show on three consecutive examina¬
tions, at intervals of a week, absence of the causative
organism.
8. All patients on discharge from hospital should be warned of
the danger they constitute to others. Any infringement of
the rules, as to abstaining from the preparation, etc., of
food for others, must be made punishable.
9. Persons engaged in the manufacture, purveyance, or sale
of food or drink, must, if called upon to do so by the
health officer, furnish evidence that he or she is not a
carrier. For this purpose he or she must allow examina¬
tions of the excreta to be made. If the results are negative
a certificate is to be given to that effect which will hold
good for a certain time, unless there are definite reasons
to suspect that the person is spreading infection before the
time for the next certificate becomes due, when an inter¬
mediate examination may be called for.
10. All such examinations must be carried out free of charge.
ADDENDUM: Received. 14 April, 1915
Since the above was written, eighty more cultivations have been
made; from two of these the Bacillus typhosus was obtained in pure
culture. Thus, from the total of 200 there have been six cases from
which the bacillus has been isolated, apart from those subjects who
270
showed post-mortem signs of enteric fever. Of these two, one was a
patient dying of dysentery, the other of tuberculosis of the lungs and
pleurae. In addition, therefore, to the usually recognised carriers,
who have definitely passed through an attack of typhoid fever, I
have found 3 per cent, of persons dying from some other affection
than typhoid, from whom no history was obtained of having suffered
from the disease, nevertheless harbouring the bacillus in their gall¬
bladders at the time of death. H. H. S.
REFERENCES
Bail, O. (1901). Untersuchungen iiber die Agglutination von Typhusbakterien. Prager
Med. Wocbenscbr., Nos. 7. and 12.
Besan^on, F. (1910). Precis de Microbiologic clinique, p. 256. Masson et Cie, Paris.
Blachstein, A. G., and Welch (1891). Johns Hopkins Hosp. Bull. II, p. 96.
Browning, C. H., Gilmour, W., and Mackie, T. J. ( 1913 ). The Isolation of Typhoid bacilli
from faeces by means of brilliant green in fluid medium. Journ. of Hygiene, XIII,
PP* 335-34^.
BrOckner (1910). Ueber Nachuntersuchungen bei Personen, die vor Jahren Typhus durch-
gemacht haben. Arb. a. d. Kais. Gesundbeitsamt , XXXIII, pp. 436-442.
Bruns, H., and Kayser, H. (1903). Ueber die Verwertbarkeit des Agglutinationsphanomcns
zur klinischen Diagnose und zur Identiflzierung von Bakterien der Typhus-Coligruppe
(Paratyphus u.s.w.). Zeitscbr. /. Hyg., XLIII, pp. 401-425.
Busse, O. (1908). Ueber das Vorkommen von Typhusbazillen im Blute von nicht Typhus-
kranke Personen. Munch. Med. Wocbenscbr., LV, pp. 1113-1116.
Chiarolanza, R. (1909). Experimentelle Untersuchungen iiber die Beziehungen der Typhus¬
bazillen zu der Gallenblase und den Gallenwegen. Zeitscbr. f. Hyg., LXII, pp. 11-32.
De Grandmaison (1900). Arcb. de med. expSr. et <Panat. patbol ., p. 289.
Doerr, R. (1905). Experimentelle Untersuchungen iiber das Fortwuchem von Typhusbacillen
in der Gallenblase. Centralbl. f. Bakt., XXXIX, pp. 624-634.
Droba, S. (1899). Der Zusammenhang zwischen Typhusinfection und Cholelithiasis auf
grund eines in der Klinik operierten Falles. Wien. Klin. Wocbenscbr ., XII, pp. 1141-1145.
Dupr£, Ramond and Faitout. Quoted by Bezan^on, Precis de Microbiologie clinique, p. 255.
Eisenberg, P., and Volk, R. (1902). Untersuchungen iiber die Agglutination. Zeitscbr.
f. Hyg., XL, pp. 155-195.
Faitout, F., and Ramond, P. (1896). Angiocholecystite a bacille d'Eberth. Compt. Rend.
Soc. Biol., pp. 1130-1132.
Findlay, J. W., and Buchanan, R. M. (1906). Case of Typhoidal Cholecystitis in which the
usual symptoms of typhoid were absent and in which the bacillus typhosus was isolated
during life from the cystic and intestinal contents. Glasgow Med. Journ., LXV,
pp. 177 - 193 *
Fischer (1909). Klin. Jabrb., XXII.
Fornet, W. (1912). Statistisches iiber den Typhus und die Typhusbekampfung im Sudwesten
des Reichs. Arb. a. d. Kais. Gesundbeitsamt, XLI, pp. 448-502.
Forster, J., and Kayser, H. (1905). Ueber das Vorkommen von Typhusbazillen in der Galle
von Typhuskranken und 4 Typhusbazillentragem.’ Munch. Med. Wocbenscbr., LII,
pp. 1474 - 1477 *
271
Fxoick (1908). Klin. Jabrb., XIX.
Hamilton, A. (1903). Journ. American Med. Assoc., XL, p. 576.
Hewlett, R. T. (1908). Manual of Bacteriology, 3rd Edition, p. 335.
Hilgermann (1909). Kin. Jabrb., XXI.
Hiss, P. H., and Zinsser, H. (1910). Textbook of Bacteriology, p. 411.
Hunner-Writer (1905). Wien. Klin. Wocbenscbr., p. 1476.
Kayser, H. (1907). Ueber Untersuchungen bei Personen die vor Jahren Typhus durchgemacht
haben und die Gefsihrlichkeit von 4 Bazillentragem.’ Arb. a. d. Kais. Gesundbeitsamt,
XXV, pp. 223-228.
Kirstein, F. (1904). Ueber Beeinflussung der Agglutinerbarkeit von Bakterien, inbesondere
von Typhusbacillen. Zeitscbr. f. Hyg., XLVI, pp. 229-260.
Koch, J. (1909). Typhusbazillen und Gallenblase. Zeitscbr. f. Hyg., LXII, pp. 1-10.
Krokiewicz, A. (1908). Typhusbazillen im Blute und Gruber-Widalsche Reaktion bei Pyamie.
Wien. Klin. Wocbenscbr., XXI, pp. 1633-1634.
Lehmann, K. B., and Neumann, R. O. (1912). Lehrbuch der speziellen bakteriologischen
Diagnostik, Part II, 5th edit., p. 324.
Lentz (1905)- Klin. Jabrb., XIV, p. 475.
Lipschutz, B. (1904). Ueber die bakteriologische Diagnose des Typhus abdominalis. Mit
Hilfe des v. Drigalsld-Conradischen Nahrbodens und der Agglutination. Centralbl.
f. Bakt., XXXV, pp. 798-811.
Lumsden, L. L., and Anderson, J. F. (1911). Report No. 4, on the Origin and Prevalence
of Typhoid Fever in the District of Columbia (1909-1910). Bull. 78, Public Health
and Marine Hospital Service of United States, Washington, pp. 1-196.
Marmann, J. (1908). Bericht iiber die Tatigkeit des bakteriologischen Untersuchungsamts
zu Gottingen im Jahre 1907-08. Hyg. Rundschau, XVIII, pp. 1013-1020.
Meyerstein, W. (1907). Ueber die bakteriologische Bedeutung der Gallensalze. Centralbl.
f. Bakt., XLIV, pp. 434-4+0.
Muller, P. Th. (1903). Ueber die Immunisierung des Typhus bacillus gegen spezifische
Agglutinine. Munch. Med. Wocbenscbr ., L, pp. 56-61.
Nicolle, C., and Trenel, M. (1902). Recherches sur le phfcnomine de Pagglutination.
Ann. Inst. Pasteur, XVI, pp. 562-586.
Nieter, A., and Liepmann, H. (1906). Ueber bemerkenswerte Befunde bei Untersuchungen
auf das Vorhandensein von Typhus bazillentragem in einer Irrenanstalt. Munch. Med.
Wocbenscbr., LIII, pp. 1611-1612.
Nieter, A. (1907). Ueber das Vorkommen und die Bedeutung von Typhusbazillentragem in
Irrenanstalten. Munch. Med. Wocbenscbr., LIV, pp. 1622-1624.
Pies, W. (1907). Untersuchungen iiber die Wachstumsgeschwindigkeit der Typhusbazillen
in Galle. Arcbiv. f. Hyg., LXII, pp. 107-127.
Rosenau, M. J., Lumsden, L. L., and Kastle, J. H. (1909). Report No. 3, on the Origin
and Prevalence of Typhoid Fever in the District of Columbia (1908). Bull. 52, Public
Health and Marine Hospital Service of United States, Washington.
Scott, H. H. (1913). Widal’s Agglutination Reaction as an aid in prognosis in Enteric Fever,
based on a study of over 1,500 consecutive blood examinations. Practitioner , Nov. 1913,
pp. 589-608.
Scott, H. H. (1911-1913). Report of Bacteriological Department, Annual Report Jamaica
Medical Service, 1911-12. Also Annual Report, 1912-1913.
Scott, H. H. (1912). Report to the Tropical Research Committee, pp. 30 et seq.
Stewart, J. (1901). A Report on 620 Cases of Typhoid Fever. Brit. Med. Journ., I,
pp. 1463-1467.
Totsuka, K. (1903). Studien iiber Bacterium coli. Zeitscbr . /. Hyg., XLV, pp. 115-124.
Wassermann, A. von (1902). Ueber Agglutinine und Pracipitine. Zeitscbr. f. Hyg., XLII,
pp. 267-292.
Zinsser, H. (1908). A Case of Typhoid bacilli in the Gall Bladder. Proc. of New York Pathol.
Soc ., pp. 78-81.
Table II.—Post-mortem findings and results of cultivation of the Bile from 120 consecutive
272
.5
* to
a*
£
M
to
u
3
•5
o
*
8 .
-a
c
d
o
■S
•J
CJ
T3 , 6
§ £ £
* S
" £
<-
-3
*a
a
o
I I
I I
I I
T3
9
a
c
W
o
vO
•§ §
* i
o
•a o
w to
^ n
o N
-2d £
It
3
T 3
C
-o
ja
•e
e
w
-o
C
TS
I
•a
o
£ £
B
e
£
a
w
*0
*5
c
2
1
3
u
•a
8
<
* £
•a
o
C
8 .
C
cS
•a
M
w
£
T 3
B
Yes
No
No
(0
£
M
V
><
No
No
O
Yes
Yes
No
No
No
0
%
0 J
Z >
«o
>«
No
No
No
No
0
£
No
No
0
55
No
No
No
No
No
0 £
Z >-
00
£
00
CO
N
N
§•
CO
_ N
^ N
to
c«
co
OO
H
| 5*
| *9
x
s'
s'
s
s’
s'
s'
s
fa
s
S
fa
i * 1
cJ
cj
d
32
h
<
d
fa
^ t/3
£
fa
u.
in
</i 0
R.
w
<
S
<
J
cj
►— >
6
X
<
d
<
►0
N <
-
H
co
4 -
to
vO
N
00
O'
0
=
12
x 3
to 0
j Dysentery Large intestine gangrenous, — Shiga type
| mitral vegetations, renal in-
I farcti
273
CJ
«
a
>%
H
I I
I I
I I I I
I I
I I
I + + I
I I
I I
I I
i ii
i n
U
2
9
CU
O
a
•c
€
2*5
§1
£*c
a. w
fl a.
*S>
*C
■tf
*C
M ,
S.
2
•c
&
no
9
«•
•a
•e
2
(3
'I
|
'!
*T3
nO
*>
■8
*o
V
•X3
V
« S o
■8 a I £
1
no
V
1*
*o
c
1
g
g
g
g
g -if S
g
g
1:s -s
g
•a
•a
(fi
C
VC
c
•a
^ 111
vd
C
cC
d
|I 3
id
d
o
o
o
O
o
U
o
o
3
3 6 2
o
o
3
H £
3
C
G
-a s
3 §
£ 25
£
*e
*2 *•
S' 9
•8 c
« .a o
s a *c
~ 8 2
o m e
25 0 «
*3
•n
M
U ft.
> 4>
v 0
•M
*2
a
a |
? g
a s
*§ *fi
a
a
(A
a
■5
a
-T3
2
u
•g M
s
V
J
V rt >
*T 3 r. qc
rt . 2 *a C
6 « u *3
S 4 1 1
£ sS A >
« >
2 u
v *C
d 5
O c
2 &
$
*+
fx*
y
3
£
a • r
J s-1
« I
5 u
°° SL
*rs
•s
o o
£
o
o o
£
o o Z
£ 25 5H
o © 5
£ 25 >«
oooo o o S o ojjo
2: 25 >« £ £ >• £
OOOO OOajO bj O
fc £ fc £ £ fc >« £ >« >. 25
2 -
U* ft.
£ Q
W c /3
OQ
J
55
w
u
P 5
ft.
w
O
J
* a s a
< d ft.*
* ^ ei £
a a w >
^ a
w £ PQ
o - m
O' o ~
i Phthisis | Liver abscesses, gall-stone
l ulcerated into duodenum
2 74
1
w
J 5
o
c
0
be C fl
<•" SJL
I I I
+ 111
I I
*C
ex.
8
i i
B 0 •
. a g
■8 s §
U 4i P
a
;c
-3
1
ex
I i
O s 2
■c
C
w
•o
e
i i
i i
i + +
•a
3 o
s
•1
t:
•a
I
■a
4 'S
i e
•a ?
•3
U
s
•c
*i
§4
n
V T3
J §
3 *
*2 ;
3
c
c
3
1
“> n C
4-» 4/ O
> ** *rt
»*« c s
•a
&
a
8 .
•c
>
e
4J
X
W
‘C
s
c
w
•a
‘6b
u
C
&
c
o
M U *3
* S > S
JS 2
•c A
, c £
H w «/>
•a
tc
t> o O c*
>< £ £ ><
fa S fa s
55 z z, Z >«
tc-o „• *
’ELS
2 S t S
o s,
5* 55 55
o Z o
£ > Z
55 55
55 >
o
S 5
o S o
Z > Z
% s
N W O
fa
fa
u
u
«
u
>
b/2 H
< d
o
w
03 E/2
S *
P 5 <
fa
<
5 fa s
s
Sad
< > d
$
tr> VO OC
Table II. — continued.
275
l g i g g
tc <c <c
c c c c c
00000
u u u u u
3
Table II. — continued.
276
0
£
0 l>
0
0
0
Id
at
0
1
|
|
1
1
I
1
1
1
0
0
1
1
1
1
B
O
ex' 73
£§
"5
a.
2
"5
v»
*0
u
3
U
+J>
c/)
ai
</J
fti
fti
M
•-)
m
3
O
x
1
+
1
+
1
1
1
1
1
1
1
!
1
1
1
1
1
1
Ou
H
«
3
O
rt
2
C
O
3
*3
a
u
0
£
1
0
c
f
i
ialignant endocarditis,
embolic pyaemia
T3
9
&
c
3
«
C
CL
In
n
Xi
c
2
3
a.
Sv
Cl
c
a
0
*3
S.
ifx
s.
-a
e
"O
•d
•d
T3
1
T3
i
*d
*d
r 40
2 u
•§ a
•T
•s
0
•d
•0
3
•6
•c
a e
0 s
•d
0
Pi
0
g
O
g
i
i
0D
1
44
g
O
g
3 Ja
•8 a
1
V
g
•a
On
•»
*3
•ft
*2
S'
•ll
0
2
VC
c
0
VC
C
O
VC
c
0
1
•tf
iC
0
*0
2
44
VC
c
0
*a
0
JS
3
VC
c
0
&
a
a
4 -*
.c
•|
i
a
M +*
fc.S
<c
c
0
I
u
u
0
u
(J
0
u
u
H
h
u
j
Pn
O
U
u
0
<
O
V
"O
•0
•0
•a
a
_
S
E
;§.
M
CL
3
;s
2
O
t>0
.9
cfl
■3
'd
§
l
1
u
4-1
IX
8
In
►
O
0
2
1
V
0
V
ri
•3
2
n
2
t)
«
Cl
0
0
V
>
V
V
V
>
too
J4
'a
M
O
CU
•O
*d
ct
C
O
’•3
-d
«
’V
1 4
•d
cs
rt
1
•d
d
•4N
y
•8
0
*c
d
U*
•e
•a
M
3
3
£
•g
3
£
3
|
6
3
•c
3
2
&
£
<
J8
4J
4-1
4-1
4-1
2
44
4-1
4-4
c
c 1
3
Pd
C
w
0
0
O
55
1
§
O
55
O
55
O
55
O
!5
0
55
c
P4
0
55
pj
/V.
« 1
0
T3
+
1
1
:
1
:
1
1
:
ev.
1
1
1
Ctf
M
>
w -
Pt-i
*2 ■ £
0
O
a
Four
post
morti
si
V
>
£
55
s2
5?
55
55
55
55
55
55
55
£
55
55
j2
w
i «
O
t)
0
0
0
0
0
0
0
0
•ft
o>
0
00
0
0
•
Cl
0
s
0
2 0
“ &
55
>
55
55
>«
55
55
55
55
55
>
55
>*
z
><
55
>
z
w £
bCfl (1
N
M
N
N
%
Os
0
/In*
9-
Os
0
US
0
m
Os
m
N
tv
ov
us
N
OO
m
»s
tv
ib
Sex
pi
S
s
s
pi
S
S
Pi
pi
S
pi
pi
pi
a
S
a
pi
s
I
:
-a
z
O
■3
CP
pi
0
s
<
S. Y.
Q
C/5
u
<
J
O
PQ
►*n
in
*-—>
in
_i
P5
►A
M. W
pi
W. A.
W. S.
pC
A. R.
W. B.
d
0
N
r*s
'+■
us
VO
tv
00
Os
0
N
m
-**
US
SO
tv
55
tv
tv
tv
tv
tv
tv
tv
tv
tv
tv
00
00
OO
00
00
00
OO
00
Enteric Fever Diagnosis
2 77
O
CL
T3
I I
C
"5 |
^ o
«i S
I I
+ i
3
~cL
I i
I I
•c
w
c
W
•as
3 g
-G O
*■8
O
1°
° i
3 v
|I
I!
S £
e o
S> 6
K 8
xS
*3 o
c* £
*1 8^
. i§> r
1 Is I lis>
"3 § C * ^ 3 -C
> U Q U
'O
o
3
*3
*3
2
'C
8 .
be
G
.G
bO
•c
i. | g u
Jo Is *g
§ 3 e
u ^ w
+ + I
c
-G
-3
G
I I
•a
j2
S *3
s § «
■fa §
H O
V >
“ e '5
-o . S
C *L o
§ is
**• g
2 o
* 8.
« C rt
<— s;
«*
2
M
£
2
a
3
TJ
2
.O
12
2 .8
2
*i3
S!
r*
*3
G
E>
2
s
.2 c
* °
2 c
J 5 £
&
u
•g
*•
c
U 3
o
*3
'£
2
'C
8 ,
2
V
G
(S
te
o
Z
*E
«■
-s
.2 £
c _
I 2
I -s
V k-
bo u
1 1
£ Q
Q -
- -
- -
-.-
—
- -
—
-
-
■ -
0
o
O
o
o
O
o
O
c
•ft
c
o
o
O
z
z
z
Z
z
z
z
Z
Z
>«
>
Z
z
Z
Z
o
o
o
o
o
c
•ft
o
o
o
o
o
o
o
o
Z
z
z
Z
z
Z
>«
z
Z
Z
Z
z
Z
Z
Jh
Z
*r\
oo
N
N
N
cl
o
VO
hv
r-v
N
ON
O'
O'
NO
N
c<->
c*")
w-k
SO
M
H
H
n
c<*v
"
"
"
"
PL
d
S
tJL
PL
s
lu
Pl
PL
s
pL
J
d
o
03
J
S
5?
J
w
Q
S
>
a
d
m
J
Q
«
•—>
s
H
<
CL
d
Q
r 1
d
X
c/i
<
►—»
W
w
s
oo
00
O'
oo
8.
ON
N
ON
r*i
O'
s
uv
ON
*
IN.
O'
8
OOI
0
O
C^i
O
Not made (moribund) ! Dysentery
Table II.— continued.
*78
U
>
-a , c
c w o
3 «« *-*
r° SL °
fa ^ g
Ul C N
-3
.2 -C C
w w
b *JS
c
S
>s
"O
£ :s
e
w
£
JC
« £
g w
a 4
•t
w
st
rt
•3
•2 (y rt
o tt: *** Sd
c o .u jii
C 1 _ Q.
3 cs jr
S 8 C i
CL fa CL
O
y
c
52
& £
!> vS S
c
x
©
52
o
x
e c
£ £
© o
£ fc
©
x
© o £ o
^ z > y
o © So
y y > y
s s s
S * S S fa S fa s'
DC
u
PC
J 3
u
O
CD £
£ S
X
fa
co
H
CD
U
PC
fa
S
£
£ fa
> u
ffi £
CO fa
cc
R. S. ...j F. 26 | No No ... ! Mitral rcgurgation Confirmed
1908
March April
Jdy Aag. Sept Oct.
Nov.
Dec.
1909
50
45
Fth.
May
Jane
0
Chart II.—Notifications of Enteric Fever cases in Kingston during 1908.
Chart III.—Notifications of En 1
28
1912
1913
Chart VI.— Notifications of Enteric Fever cases during 1912.
Chart VII.—Notifications of Ent<
282
Chart IX.—Total numbers of notifications of Enteric Fever cases in Kingston,
month by month, for the years 1908-1914.
:8 4
Chart X.—Average of notifications of Enteric Fever cases in Kingston,
month by month, for the years 1908-1914.
28s
ON THE OCCURRENCE AND PRE¬
VALENCE OF DISEASES IN BRITISH
NEW GUINEA
BY
A. BREINL
FROM THE AUSTRALIAN* INSTITUTE OF TROPICAL MEDICINE
(Received for publication 4 February , 1915)
Plates XIX—XXVI and Map
CONTENTS
Introduction
PAGE
285
Malaria
288
Filariasis
29O
Leprosy
*93
JUXTA-ARTICULAR NoDULES
*94
Curious Fevers
*97
Yaws
298
Venereal Diseases
*99
A Peculiar Disease, characterised
by Arthritis, Osteitis and Periostitis
300
Ulcers in the Tropics
304
Ulcus Tropicum
3°6
Contracting Sore
309
Ulcus interdigitale destruens
313
Gangosa
...
3*4
Skin Diseases
3*4
Concluding Remarks
3*5
Appendix I. — List of Papuan Mosquitos
3*7
References ...
3 **
INTRODUCTION
In the following pages are embodied the scientific results of two
journeys to the coastal belt of British New Guinea, which were
undertaken for the purpose of mapping out the incidence and
geographical distribution of tropical diseases among the natives.
The first journey took place during the months of July and
August, 1912, and included the coastal belt east of Port Moresby
and north of Samarai as far as the Mambare River.
286
During the second journey, lasting from the end of June to the
beginning of October, 1913, the coastal regions west of Port Moresby
as far as Daru were visited. To the Purari River the journey was
accomplished on foot, and continued from there in native canoes.
The coast of New Guinea cast of Port Moresby is fairly densely
populated, the natives living in numerous villages, ranging in size
from three or four houses to large settlements with approximately
3,000 inhabitants, although very large villages are rare.
The majority of the natives on the south-east coast live by
agriculture and fishing. The villages consist of a number of houses
sufficiently large to provide a comfortable shelter for one family.
Some of the villages are built on high piles in the sea, the houses
being connected by bridges, but most of the traffic between the houses
is carried on by native canoes.
In a great number of the villages situated further inland the
houses are built on both sides of a main street, since the Government
advises, for obvious reasons, this scheme of laying out the villages.
The majority are built on sandy soil and surrounded by more or less
dense scrub or jungle.
The coastal districts west of Port Moresby, as far as Orokolo
Bay, are of the same nature as those on the south-east coast. The
villages are densely dotted all along the coast, and in many instances
a few miles only intervene between neighbouring villages.
After passing Orokolo Bay the nature of the country undergoes
a complete change. The coast between the Purari and the Fly
Rivers is broken up into a continuous delta. There are a number of
large rivers connected by numerous cross channels and cross streams,
and the land is entirely of a muddy and swampy nature. The
Villages are built on mud flats, and the traveller may journey for
days and weeks without stepping on firm soil.
The villages there consist of houses of varying size; besides small
huts accommodating one family only, there are very large houses,
inhabited by a number of families, and large club houses where the
unmarried boys dwell.
Throughout the west the native houses are built on high piles,
and the different sections of each village are connected by bridges,
consisting of a light scaffolding with loose saplings laid across, which
are renewed at rare intervals. Still further west, on the Fly River,
287
each village consists of a small number of very large houses which
give shelter to a hundred or more people, and are partitioned off
into cubicles each occupied by a single family.
Generally speaking, the natives of the south-east and west coasts
as far as Orokolo are highly civilized, and have been in constant
contact and communication with Europeans for nearly a quarter ot
a century. West of Orokolo the natives become Ids and less
civilized. In some districts, especially in the neighbourhood of the
Kikori and Turama Rivers, numbers of villages are not under
Government control, and many of them have not even been visited
by Europeans. The villages situated in the Fly Estuary, and the
inhabitants of some of the islands, as Kiwai Island, are as highly
civilised as the natives in the neighbourhood of the capital of British
New Guinea, Port Moresby.
The medical survey was carried out with all possible thoroughness
and care, and it may be assumed that cases of almost all diseases
prevalent in New Guinea have been encountered in one village or
another.
During the journey a number of blood specimens were obtained
in order to map out the distribution of filariasis and malaria.
The less civilised the districts, the fewer natives were found
willing to supply a drop of blood for a film. Unfortunately in
several districts, especially around the Kikori River, it was
impossible even to obtain photographs of natives suffering from the
various complaints. As opportunity offered, sores were scraped with
a sharp spoon, and the granulation tissue so obtained preserved in
70 per cent, alcohol for future microscopical examination.
Speaking generally, a number of diseases such as various types
of ulcers, malaria, filariasis and elephantiasis, juxta-articular
nodules, gangosa, yaws, and several skin diseases, mainly tinea
imbricata, were encountered in most of the villages visited. It must
be borne in mind that for a considerable time frequent intercourse
and trade took place amongst the natives on the south coast between
Orokolo and Samarai. Every year the natives of Port Moresby and
several adjoining villages, who are the potters of New Guinea, travel
west in the lakatois (large sailing canoes), exchanging their pots for
sago which grows abundantly in the western parts. The journeys
must have offered ample opportunity for the spread of contagious
288
and infectious diseases. In more recent times the opening up of the
couhtry, and the drawing of the supply of native labour from
districts the inhabitants of which had hitherto not been wont to
travel, has given rise to new channels for the propagation of disease.
It is, however, noteworthy that several diseases, such as gangosa,
juxta-articular nodules, yaws and tinea imbricata, and different
types of sores, were seen in natives who were unable to travel any
considerable distance without endangering life and limb on account
of hostile neighbours.
MALARIA
With the exception of a few districts, malaria occurs with
varying severity throughout British New Guinea, although its
frequency is dependent on seasons and local conditions. New
Guinea, situated within the monsoonal belt, has two distinct seasons
—the north-west beginning at the end of November and lasting
until March or April, when land breezes prevail, and the south-east
season when the prevailing wind comes from the sea.
In the coastal villages malarial fever is more prevalent during
the north-west season when mosquitoes are in evidence. During the
south-west time, on the other hand, malaria is far less common, since
the sea breeze prevents the mosquitoes from swarming into the
villages.
In the inland villages in sheltered positions, on the whole, no
marked seasonal variation is observed. Coastal villages in exposed
positions are comparatively free from fever, whereas coastal villages
sheltered from the breeze show a large percentage of children with
enlarged and palpable spleens. In the villages west of Orokolo,
built on mud flats, which were visited during the south-west monsoon,
no cases of malarial fever amongst the natives were encountered,
although fever is said to be prevalent during the north-west season.
The natives contract malaria as a rule in early childhood, and
acquire, in the course of years, an active immunity against the type
of malarial infection prevalent in their place of residence. The
incidence of malaria in natives decreases in direct proportion to their
age.
The number of children with enlarged spleen gives a definite
289
indication of the amount of malaria in a given locality. The spleen
census was taken by determining the number of children with
enlarged spleens in the different villages, and blood films were made
of those children who showed considerable enlargement of the spleen
to ascertain the type of malaria prevalent.
On several occasions the spleen index of babies in arms was taken
separately from that of children of ages ranging between 5 and 11
years, and the former was considerably higher than the latter, in
one instance 95*1 in comparison with 58*6.
The accompanying map shows that simple and malignant tertian
types of malaria are fairly equally distributed and more common
than the quartan type; the latter is more prevalent in the furthest
north-east corner, on the mouth of the Mambare River, and is not
infrequently met with in the Mekeo district.
Out of 245 blood films taken from a small number of native
children, 132 contained malarial parasites, sometimes in considerable
numbers; at other times only a few ring forms were seen in a whole
film.
Out of 245 blood films examined, 48 contained the parasite of
malignant tertian, 34 of simple tertian, and 27 of quartan malaria.
In 23 the type of the parasite could not be determined with certainty.
The white population of British New Guinea suffers, with hardly
any exception, from occasional attacks of malarial fever, and not one
white settler was met who had been entirely free from attacks of fever
during his stay in the country.
Of Anophelinae, N yssorhynchus annulipes and Cellia punctulata
were collected, the former being the more common mosquito,
occurring in the proportion of about 10 to 1 of the latter.
The occurrence of A 1 yssorhynchus annulipes in numbers through¬
out the districts where malarial fever is most prevalent, and the fact
that the development of Plasmodium falciparum has been traced by
Kinoshita (1906) in this mosquito, make it extremely likely that
Nyssorhynchus annulipes acts as intermediary host of the malarial
parasite in New Guinea.
Of other Anophelinae, N yssorhynchus bancrofti has been
collected, but this species does not transmit the parasite.
290
FILARIASIS
Thorpe in 1896 drew attention to the fact that the microfilaria
in the blood of the natives of the Tonga Islands does not show the
same periodicity as observed elsewhere. Lynch (1905) confirmed
this statement, finding that in 105 out of 156 cases of filariasis
amongst} the Fijians, microfilariae were present in the peripheral
blood both by day and night.
Similar observations in the cases of filariasis among the natives
of the Philippine Islands led Ashbum and Craig (1907) to the
creation of a new species, Filaria philippensis. The larvae, although
resembling closely Microfilaria nocturna , do not possess any
periodicity, and occur in approximately the same numbers in the
peripheral blood during all hours of the night and day. The
movements of this microfilaria are described as progressive and
lashing, the sheath is very tightly fitting, so that the microfilaria
cannot slip backwards and forwards within it, as in the case of
Microfilaria nocturna. At the posterior end the tail diminishes
progressively and uniformly to a fine thread-like point. Bahr (1912),
Fiilleborn (1913), and many other observers, have failed to
discriminate between the larvae of Filaria philippensis and that of
Filaria bancrofti.
Fiilleborn (1911), utilising material collected in the Bismark
Archipelago and German New Guinea, pointed out that 10 to 60
per cent, of the natives showed large sheathed microfilariae in their
blood, which were of the same anatomical structure as Microfilaria
bancrofti but did not possess the typical nocturnal periodicity. More¬
over, the adult worm of the Fijian microfilaria, according to Leiper,
cannot be differentiated from Filaria bancrofti.
Fiilleborn (1912) concluded, therefore, that in the present state
of our knowledge two biological varieties of Filaria bancrofti can be
differentiated; the larvae of one variety have a pronounced nocturnal
periodicity, whereas the larvae of the other, mostly occurring in the
Pacific, are present in the peripheral blood at all hours of the day
and night.
A troop of twenty-three Samoans, who were visiting Europe, were
examined by Fiilleborn (1912). He found that nearly half of them
harboured microfilariae in their peripheral blood, morphologically
291
identical with Microfilaria nocturna , but without any marked
periodicity.
Whilst travelling through the coastal districts of British New
Guinea, thin smears were made from the day blood of a number of
men, women and children to be examined for the presence of micro¬
filariae. It was found impossible to use thick films for this work,
since previous experience had shown that they deteriorate much
quicker than thin blood smears, and after exposure for some time to
the moist tropical heat are difficult to dehaemoglobinize. On account
of the short stay in the different villages it was possible to take
blood films in the day-time only, as the blood examination of the
same natives on two occasions would have meant too great a delay.
A more thorough and extensive filarial survey will be undertaken at
a future date.
Although the number of blood examinations was comparatively
small, the results indicate that the natives living in the coastal belt
of eastern and north-eastern New Guinea harbour filarial larvae in
their blood to a greater extent than in the western parts. Out of 166
blood slides taken at random on the first journey, twenty-four
contained microfilariae, giving an infection index of slightly over
17 per cent., whereas out of 166 films taken during the second
journey, microfilariae were discovered in eight slides, being about
5 per cent. A more extensive survey, taking samples both in the day
and night time, would almost certainly reveal a much larger
percentage of infection.
On two occasions only the blood of some of the carriers was
examined, and two and three respectively, out of about fifteen,
proved to be infected with filarial larvae, which showed the typical
nocturnal periodicity.
Morphologically the microfilaria found in the blood of New
Guinea natives does not differ essentially from typical Microfilaria
nocturna. Table 1 gives the measurements of a number of micro¬
filariae, stained by Giemsa's method, and for comparison are
appended Fiilleborn’s average measurements of typical Microfilaria
nocturna and of microfilariae found in the blood of Samoans.
The experience gained in New Guinea coincides with Bahr's
observations in Fiji, and Fiilleborn's in the South Pacific, that there
exists in British New Guinea a microfilaria morphologically identical
292
Table I.—Measurements of Microfilariae found in the blood of New Guinea natives; films taken
in the daytime.
Nerve spot Excretory 1
from j pore from :
anterior end anterior end
Inner body
from
anterior end
1
Length of
inner body
j Gi cell
' from
' anterior end
| Anal pore
from
! anterior end
1
Total
j length
%
0
,0
%
0/
,0
I 0/
/o
1
! * 8-5
28*1
468
5 °
...
81*4
4<>5
2
1 9 * 1
28-5
52-3
5 °
73*8
88 -o
410
3 1
* 7*3
27-9
486
38
848
358
4 !
1 6* >
26-6
48-9
60
689
85*3
450
5 ;
20-1
* ...
57*4
35
74 * 1
...
340
6 1
> 9*3
286
45*7
7 °
73*4
35 °
7 !
1
19*6
29*4 j
40*0
60
86-8
3<>5
8
1
1 6*4
26*2
47*8
55
73*8
« 7'3
366
9
18*2
27*3
50-0
45
7**5 ;
82*9
440
1 ° j
21 *o
29-5
5 1 * 1
52
74*5
88-0
1 352
.. !
1
19-6
316
607
30
74*4
88*5
395
12 !
■S -5
3°*3
48*4
60
75*7
33 °
*3
28-0
49*6
56
87*0
3 *°
<4
i »-4
29-0
5°*4
54
7 ! ‘4
245
Average 18-7
! *8-5
49*9
5 *
73 *»
860
362
Fulleborn’s average measurements of typical Microfilaria nocturna —
iR -3
j 28*6
70*6
1
83-5 :
232*5
Average of non
•periodical Samoan Microfilaria-
-
19*3
1 1
1 285
...
| 1
70*2
82*9 |
272*5
*93
with Microfilaria nocturna , but not possessing the typical periodicity,
in addition to microfilariae which are only present in the peripheral
blood at night time.
It is interesting to note that only one specimen of Stegomyia
pseudoscutellaris was collected in Samarai on the south-east coast
of New Guinea, which species was not encountered anywhere else.
The patchy distribution of filariasis is quite as well marked in
New Guinea as in other parts of the tropics; for example, in one
village nearly 3 to 5 per cent, of the male population suffered from a
soft swelling of the groin glands, so typical in filariasis, whereas in
the neighbouring villages cases were only rarely noticed.
Cases of elephantiasis were seen in varying numbers throughout
the districts visited. Of nearly every case of elephantiasis blood
films were taken, but in no instance were microfilariae found in the
blood. The arms and legs were most frequently affected, the arms
perhaps more often than the legs. The genitalia were not so
frequently implicated.
Plate XIX, fig. 1, shows a typical and far advanced case of
elephantiasis of the scrotum, affecting at the same time one leg; and
fig. 2 shows a typical case of elephantiasis of the labium minor of the
vulva.
LEPROSY
The distribution of leprosy in New Guinea is of some interest.
This disease has already been referred to by W. MacGregor in the
yearly report for New Guinea, 1888-1889, where it is stated that
‘ Leprosy is not an uncommon disease amongst the natives, but does
not present its worst form and has not been communicated to any
white person.'
On the whole, in the districts visited, leprosy was found endemic
in only three, namely, the Trobriand Islands—a number of islands
off the north-east coast—the Mekeo district on the St. Joseph River
in the Central Division of British New Guinea, and in one district
of the Gulf division.
Only very few cases were seen on the south coast east of Port
Moresby, and not one definite case west of the Vailala River.
Altogether eighteen cases were diagnosed, four cases in the
Trobriand Islands, seven in the Mekeo district, and eleven in the
villages on the south coast between Isiu and Orokolo, west of Port
Moresby.
The three different types, namely, nodular, nerve, and mixed
leprosy, were found.
Plate XIX, fig. 3, shows one of the most advanced cases of
leprosy seen.
It is an interesting fact that the natives of the Trobriand Islands
know leprosy well, having a special native name for the disease,
namely, Kai-gwai-guia. When the natives were asked to bring
forward these cases, all produced were typical lepers.
Considering the number of natives seen, and the large area
traversed, if becomes apparent that leprosy is present in New Guinea,
although by no means prevalent. A great number of ulcers of various
descriptions resembled leprosy at the first glance, but careful
examination excluded the diagnosis.
JUXTA-ARTICULAR NODULES
The occurrence of juxta-articular nodules was first described by
MacGregor in 1901 in New Guinea. Later, in 1904, Steiner described
the occurrence of tumours amongst Javanese, appearing as hard,
round irregular nodules below the skin, and situated always in the
neighbourhood of the joints. The skin above the tumours was
moveable, and the nodules were not connected with the underlying
fascia. On histological examination these nodules consisted of
concentric layers of hard fibrous tissue, which was degenerated in
the centre into coarse, irregular, structureless masses. Various
staining methods did not reveal the presence of any bacteria.
Two years later, Jeanselme (1906) described the occurrence of
similar tumours amongst the natives of Siam. Microscopic examina¬
tion showed that they were formed of three layers—a central one
consisting of degenerated tissue, an outer zone of inflammatory
reaction, and an intermediate or transitional zone.
Gros (1907) observed the occurrence of similar tumours amongst
the Algerian natives in small numbers only; ten cases were seen out
of approximately 12,000 patients. Histologically they were of
similar structure to those examined by Jeanselme.
The occurrence of the same disease was reported later by Neveux
2 95
(1907) amongst the natives of Senegambia, and Fontoynont and
Carougeau (1908) made an extensive study of this complaint in
Madagascar. In their experience, juxta-articular nodules were
found in persons of both sexes and of all ages, though the male sex
was more frequently attacked by the disease. Special attention was
drawn to the symmetrical occurrence of these nodules. They
pointed out that the seat of predilection of the tumours was the outer
surface of the extremities, especially the extensor surfaces, also, that
they occurred in groups in the skin in the neighbourhood of joints,
and especially where bone lies immediately below the skin, as near
the elbow and large trochanter.
The development of the juxta-articular nodules is very slow,
extending over years, until finally they become fibrous or fibro¬
cartilaginous. Fontoynont and Carougeau found in early nodules,
amongst the soft caseous central mass, very small white granules,
consisting of mycelium and filaments of a fungus, which they
described and termed Discomyces carougeaui .
Inoculation of this fungus into monkeys, rabbits, and guinea-
pigs, gave no results.
Lebouef (1911) observed four cases of this disease in New
Caledonia. According to Joyeux (1913) similar tumours were
frequently seen in natives of French Guiana, though the microscopical
picture differed in some essential points from that given by
Fontoynont and Carougeau. These tumours consisted of a ground
substance of fibrous tissue contained within inflammatory foci,
mostly around the blood-vessels. The absence of bacteria in the
sections caused Joyeux to consider juxta-articular nodules as a
clinical syndrome resulting from widely different causes.
Juxta-articular nodules are of frequent occurrence throughout the
coastal districts of British New Guinea. In a great number of
villages many cases were seen, only a few in others. The clinical
manifestation did not differ from that described by previous
observers. Fibrous tumours of varying size, from a pigeon’s to a
hen’s egg, occurred in different parts of the body, mainly in the
neighbourhood of joints and most frequently near the ankle joints
(fig. 4) above the large trochanter and over the olecranon (Plate XX,
ftg- 5 )-
The subcutaneous tumours were round or oval, the skin above
296
freely moveable, and they were not connected with the underlying
muscle.
They were of hard consistence, having as a rule a smooth, rarely
an irregular, surface, and occurred in the majority of cases
bi-symmetrically. In no cases were fistulous openings observed.
The majority of the affected natives were middle-aged men, only
rarely women. One young girl, about 16 years of age, showed well-
developed nodules over both elbow joints.
In some villages situated in the eastern end of British New
Guinea as many as 7 to 10 per cent, of the whole population were
affected. In other villages closely adjoining even the most pains¬
taking search did not reveal the existence of a single case.
These tumours never caused the natives any pain or inconvenience,
and it was only after a great deal of persuasion that permission was
given to extirpate one of these tumours for histological examination.
Further material was received through the kindness of Dr. Giblin
in Samarai.*
Macroscopically, in cross sections, the nodule does not differ in
any respect from a subcutaneous fibroma; it consists of strands of
connective tissue, concentrically arranged, the skin above being not
connected with the tumour, which is apparently not encapsulated.
Here and there are small irregular areas of yellowish appearance,
showing softening.
On microscopical examination of three nodules, the three zones
described by the previous observers could not be distinguished.
One of the nodules, which was extirpated with the overlapping
skin, showed that the nodule itself was well defined from the
surrounding subcutaneous tissue, which was normal except for the
presence of a certain amount of small-celled infiltration and
hyperaemia.
The nodule itself consisted of dense concentric layers of
connective tissue cells, possessing large irregular nuclei and cyto¬
plasm with fibrillar-like processes, which anastomosed and formed
an open meshwork. Here and there, irregularly distributed
throughout the nodule, were small areas where the cell nuclei had
disappeared, the tissue still showing the original texture. Numbers
• Dr. Giblin ha» informed me since that he had treated one or two Europeans suffering from
juxta-articular nodules.
297
of leucocytes were scattered throughout, but were most numerous in
the degenerated parts. On the whole, the tissue of the nodules was
well vascularised, and there was small-celled infiltration around the
blood vessels.
The bacteriological examination of the sections did not show any
microorganisms, and none of the nodules showed the ‘ Dtscomyces
carougeaui * It is probable that the fungus may only be present in
the early stages, and that the specimens obtained were of cases of
long duration.
CURIOUS FEVERS
The short stay in the different villages did not afford much
opportunity for studying the different types of fevers prevalent.
Four cases of a curious and apparently specific fever were
observed in villages on the south-east coast. All four cases showed
the same clinical symptoms, which were sufficiently pronounced to
suggest a specific fever.
According to an account given by an employer of native labour,
the disease sets in gradually, with an irregular fever. The first
symptom noticeable is an icteric discoloration of the mucous
membranes and of the sclerae, accompanied by pain in the splenic
region. The patients suffer from more or less frequent attacks of
continuous high fever, which may last for a few days or even a week,
the temperature rising to 105° F. They emaciate considerably.
The disease runs a chronic course—one of the patients had been ill
for nearly two and a half years—improvement and relapses following
each other at irregular intervals. The patients die finally with
symptoms of great emaciation and pronounced icterus.
The examination of the four cases showed a slight enlargement
of the spleen and a very marked enlargement of the liver, which in
one case extended to five finger breadths below the ribs; the sclerae
and the mucous membranes were of a dark yellowish colour. The
blood was decidedly anaemic.
The disease is well known to the natives, and is called ‘ Gobora *
in the Mailu district, and ‘ Tebi* further east.
Blood films were taken of each of the cases. Two of them showed
a slight malaria infection, whilst in the others no malaria parasites
could be detected. No other parasites were seen in any of the blood
films.
298
YAWS
Throughout the coastal districts of British New Guinea yaws is
widely distributed, and barely a single village was found entirely
free from the disease. In some districts yaws is more prevalent than
in others.
In 1897 Sir William MacGregor drew attention to the fact that
‘Yaws is universally endemic, but it is greatly milder than it is in
the Pacific, and is in fact not much taken note of. There does not
even appear to be a native name for it in any dialect known. No
European appears to have contracted this disease in the Possession. 1
The majority of the sufferers were children, and all the typical
lesions of primary and secondary yaws were seen at one time or
another. It was noteworthy that in different districts one or the
other of the clinical types of yaws prevailed. In the village of
Orokolo, for example, many of the children examined suffered from
the typical granulomatous eruption around the anus, whereas in
other districts hardly one case with this manifestation was seen
amongst fifty children suffering from yaws.
In the western parts sporadic cases of a generalized eruption of
secondary yaws were observed. The sufferers were mostly children,
and clearly showed that secondary yaws might lead to considerable
destruction of the skin. In one case, a young girl (figs. 6 and 7),
the eruption had implicated the nose, both cheeks, lip, the upper
arm and elbow, and had formed a semicircle on chest and back.
The arm was contracted at the elbow joint. On the affected parts
prominent yellowish scales were seen, which covered ulcers, roundish
in the early stages and later confluent. The fundus was formed by
raised reddish granulation tissue. In between these scabs was, here
and there, newly formed glossy skin.
In another case (fig. 8), the nose was partly destroyed by the
specific granuloma, which spread from the nose to the cheek.
Yaws was as prevalent amongst the civilised natives as amongst
those who had hardly come under the Government influence, and
had had no intercommunication with the neighbouring tribes.
VENEREAL DISEASES
It is very difficult indeed to map out the distribution of venereal
disease amongst natives during a journey from village to village.
Natives are prone to hide venereal disease from each other, and
systematic inspection, especially of women, is practically impossible.
The diagnosis of ‘ Syphilis 9 in a native community is more difficult
than in Europeans, on account of the occurrence of various ulcers
and after effects of ulcers, which closely resemble the syphilitic
lesions. Many of the manifestations of the tertiary or late stage of
yaws, as described by Castellani, are identical with those of tertiary
syphilis.
From the limited observations made, syphilis has not spread to
any extent amongst the natives in the coastal districts of British
New Guinea. This is in all probability due to the strict measures
adopted by the Government against its spread. Lock hospitals
have been established in different districts, and all cases reported are
interned and only discharged after thorough treatment.
A scanty number of isolated cases were found here and there,
but only in those districts where the natives had come in close contact
with Europeans.
ULCERATIVE GRANULOMA
A few cases of ulcerative granuloma were seen in the eastern
parts of New Guinea, several of them under the care of Dr. Giblin
in the Lock Hospital at Samarai. Only three typical cases were
encountered in natives living in villages on the east coast, and a few
sporadic cases in the western districts.
GONORRHOEA
Cases of gonorrhoea may be found in all localities where the
natives have come in contact with Europeans. In several villages in
the Goaribari district, where communism is still existent, from a
third to a half of the population were suffering from the typical
lesions due to Gonococcus. The clinical symptoms did not differ in
severity from those observed in a temperate climate, although
venereal sores were more frequent on account of the lack of personal
cleanliness.
300
A PECULIAR DISEASE CHARACTERISED BY ARTHRITIS,
OSTEITIS AND PERIOSTITIS AMONGST NEW GUINEA
NATIVES
In his article on skin disease in the tropics, Plehn (1914) mentions
that Castellani, Howard, Schiiffner and others have attributed joint
affections and periostitis to yaws, regarding them as its tertiary
stage. He also states that he himself observed similar bone lesions
in Cameroon natives, which he considered as the final result of
chronic osteomyelitis. Skiagrams, appended to the publication,
show thickening of the periosteum of the bones of arm and hand in
children.
A disease characterised by similar symptoms was encountered
amongst New Guinea natives of different ages, of whom, in several
instances, fairly reliable histories could be obtained.
The distribution of these cases was irregular, many villages being
quite free, whilst others furnished six or more cases, most of which
failed to show any signs of previous attacks of yaws. No evidence
could be gathered as to a connection between the incidence of these
two diseases.
It would be premature to state or deny definitely any relationship
between yaws and the disease in question, until cases have been
observed, where the two merge into each other without any prolonged
interval. The irregular distribution, the similarity of the clinical
histories, its absence from tropical regions where framboesia is very
prevalent, are points in favour of the supposition that the two
complaints are separate clinical entities.
This disease is exceedingly chronic, and is termed by the natives
of the central district ‘ Roaki,* and by the natives of Kiwai Island
‘ Buuo* or ‘Auma.’
The most prominent symptom is an affection of one or more of
the joints, ranging from a hot painful swelling to suppuration and
the formation of sores with fistulae in the skin above the joints,
which discharge an amber-coloured clear liquid containing floccular
particles. At the same time spindle-shaped thickenings of the larger
and smaller bones of the extremities become noticeable, which,
especially when the small bones of the hand and foot are implicated,
may lead to softening and resorption of the bone. Similar sores to
3oi
those above the joints may make their appearance above the
swellings of the bone.
The disease affects natives of all ages, men, women and children,
although the majority were young persons. The youngest patient
was about eight years of age.
As far as could be ascertained, the disease begins with an
affection of one or more joints, without any special predilection being
noticed. The joints swell up and become painful. After a varying
period the swelling may abate or increase, and in this case the pain
prevents the patient from using the affected extremity. After a
prolonged period fistulous openings may appear in the neighbour¬
hood of the joints secreting at first flocculent pus, and later on an
amber-coloured clear fluid.
The foot so affected (Plate XXI, fig. 9) closely resembles Madura
foot without the presence of the typical granules in the pus.
In other cases the bones may become affected, at first either in
close proximity to the joint or the diaphysis of the long bones, mostly
the tibia, ulna or radius; or in still other cases the metatarsal and
metaphalangeal bones may show the change. The bone thickens,
and a spindle-shaped swelling becomes noticeable which is not
painful on pressure. This thickening is so well marked that it can
readily be noticed on inspection (see figs. 10, 11).
These swellings of the various bones may remain unaltered for
years, even for the rest of the patient’s life-time. In other cases,
especially when the smaller bones of hand and foot are the seat of
the swelling, the lesion clinically resembles osteomyelitis. Fistulous
openings appear which discharge copiously at first, and in time the
whole of the affected bone may become resorbed. This process leads
to peculiar disfigurement of the feet and hands (fig. 12). In cases
where one of the metacarpal or metatarsal bones is resorbed, the
fingers or toes become either retracted or even displaced, as in
Plate XXII, fig. 13, where the large toe is in an abducted and hyper-
extended position.
The frequent occurrence of the swelling of the radial, ulnar and
tibial bones simultaneously with swellings of the metacarpi or meta¬
tarsi and swellings of the joints, appeared to furnish proof that all
these lesions were due to the same disease.
The lesions of the hand and feet referred to may easily give rise
302
to a mistaken diagnosis. More than one case on superficial
examination resembled nerve leprosy. A thorough examination,
however, excluded this diagnosis, since in every case the finger tips
were normal, the nails still well formed, and the thickening of the
main nerve trunks was absent.
In a later stage ulcers are formed in the skin over the affected
joints and bones. These are superficial, of varying size up to
25 cm. in length, oblong in shape with irregular edges; the granula¬
tion tissue is only slightly raised, of dark reddish colour, discharging
an amber-coloured clear fluid. On the whole they show a tendency
to spontaneous healing, forming dense scar tissue (fig. 14).
It was impossible to obtain any conclusive evidence as to the
sequence of the lesions of bone and skin, since the information
obtained from intelligent natives varied considerably. Some of them
pointed out that the bone and skin lesions arose independently,
others, however, stated definitely that the bone lesion preceded the
skin lesion.
Several of the cases resembled, clinically, osteomyelitis with
fistulous openings. In the majority, however, an inserted probe
did not reach the bone.
The histological examination of the granulation tissue of one case
did not show any peculiarities. It resembled, microscopically, typical
granulation tissue containing a number of phagocytes and numerous
bacteria and cocci on the surface.
Short case histories so far as obtained, and a clinical description
of a few typical cases, may be helpful to illustrate this interesting
and somewhat obscure disease.
Case /. Idiri—Woman about 35 years of age.
The patient had been ill for a long time, perhaps five years. She
never suffered from any painful joint lesions. The first symptom
noticed was a hard swelling of the tibia. A short time afterwards
sores formed above the swelling of the bones, which healed up after
a long time. The metatarsus of the big toe of her right foot began
to swell, and afterwards small openings appeared in the skin above
the swelling, discharging, for a long time, thick matter. A
considerable time afterwards these fistulae ceased to discharge, and
healed up with displacement of the large toe.
303
At the time of examination the shaft of the right tibia showed a
considerable spindle-shaped swelling. Numerous scars had formed
in the skin above and on the dorsal side of the foot. The metatarsus
of the big toe had disappeared, and the toe consequently was
abducted and hyperextended. The right wrist joint was swollen
and painful, the ulna showed in the middle of its shaft a marked
spindle-shaped swelling, about io cm. long.
Case IL Boy about 12 years of age. Comp. fig. 11.
The boy had been ill since childhood, and the disease had begun
with painful swellings of the bone. Some time afterwards large,
slowly healing sores formed in the skin above the swellings.
The boy was very emaciated. Both tibiae were curved forward,
the bone thickened and the skin above showed ulceration. Both
ulnar bones had, in their upper third, a spindle-shaped thickening
with a fistulous opening, on one side discharging liquid pus. There
was extensive ulceration on both the arms, where roundish irregular
islets of reddish, raised granulation tissue were surrounded by very
thin newly-formed skin. Both legs were ankylosed at the knee
joint.
Case III . Small boy about 8 years of age.
The patient was hale and hearty till about one year ago, when
his right ankle joint began to swell up. Shortly afterwards his right
knee joint became painful and swollen, and later the left knee and
both wrist joints became affected in the same way. When examined
the boy was considerably emaciated, both the knee and wrist joints
were swollen and painful. The muscles of both calves as well as the
muscles of both forearms were atrophic. The distal end of both of
the radial bones was considerably enlarged.
The patient did not show any sign of present or past infection
with yaws, and throughout the village not a single case of yaws
was seen.
Case IV. Man about 35 years of age.
According to information obtained from the patient, the disease
had started about twenty years ago with pains in his back and in
the joints of his arms and legs.
When examined his right foot was very much swollen, and all
304
the toes of his foot had dropped off as the result of ulceration. The
skin of the foot was uneven, lumpy, and numerous fistulous openings
were seen, some apparently closed, others secreting on pressure
whitish floccular masses. The muscles of the leg itself were atrophic,
and the skin above the much thickened tibiae covered with old scars.
The left tibia showed the same changes, simulating a chronic
oestomyelitis. The left elbow joint was swollen, hot and painful,
and the tip of the elbow the seat of a small ulceration.
The fingers of the left hand were deformed, the metacarpo¬
phalangeal joint of the fourth and fifth finger were ankylosed. The
shafts of both radial bones showed a spindle-shaped thickening, and
there were deep scars over both radio-carpal joints.
ULCERS IN THE TROPICS
A great variety of superficial and phagedaenic ulcers, apparently
of the most varied etiologies, are collected under the name 4 Tropical
Ulcer.’ The clinical differentiation of ulcers in the tropics becomes
still more involved on taking into consideration the careless habits
and the lack of cleanliness amongst native races in general. Any
superficial skin abrasion on hand or leg, which if at all cared for
would heal, may give rise to a small ulcer which on account of the
continuous irritation caused by sand, dirt and flies may spread. It
is probable that many of the large phagedaenic ulcers are not due to
one specific microorganism, but are caused by a variety of micro¬
organisms invading a small wound due to an accident.
The observations made in British New Guinea appear to bear
out this statement. On the whole the incidence of ulcers depends
largely on the state of cleanliness, which is greatly influenced by the
natural conditions and surroundings of the different villages. In
the districts west of Orokolo, where the villages are built on high
piles on muddy grounds, and where fresh water is very scarce,
ulcers of varying types are encountered in great numbers among the
natives.
The natives in these districts, where sago palms grow in great
abundance, use sago as their staple article of diet, and spend, in
consequence, much of their time in preparing sago from the trunk
of the palm. The sago palm carries large thorns on the rib of the
305
leaves and on the trunk, and the natives are often injured by them
when felling the trees. The wounds in the majority of cases are not
cared for, and undoubtedly many ulcers are originally due to wounds
caused by these thorns.
The number of inhabitants with ulcers in villages built on dry,
sandy soil is much smaller.
The hypothesis that many of these ulcers are caused through
secondary contamination of smaller or larger injuries of the skin
receives additional support from the experience gained in their treat¬
ment. A great number of the sores heal up readily under the
application of antiseptic powders only, without any specific
treatment. It must, however, be, kept in mind that the scar,
covered by a newly-formed skin on the original site of the ulcer, is
always more liable to injury and may again become the seat of a
similar ulcer.
The question of the differentiation and diagnosis of ulcers
becomes more complicated by the facts that in the majority of the
natives the ulcers are in an advanced stage, that it is impossible to
obtain a definite and reliable history, and that advanced ulcers of
many different etiologies are similar. The question of a tertiary
form of yaws is still under discussion, and the clinical symptoms
attributed by various observers to tertiary yaws may be due to other
factors.
The diagnosis of syphilis is not less complicated, and a traveller
with little experience of natives in their original state might make
the diagnosis of syphilis where its occurrence may be excluded with
certainty.
Gangosa, so similar to tertiary syphilis in its advanced stages,
causes ulcers of arms and legs which, when far advanced, are in no
way characteristic.
Many other complaints, such as leprosy, various skin diseases,
etc., may take an atypical form in natives and make a differential
diagnosis in the late stages impossible.
Histological examination of the granulation tissue does not
permit a definite differentiation of the ulcers, since the microscopic
appearance of sections of different and clinically apparently well
characterised sores is similar. Smears taken from the surface are,
as a rule, a pell-mell of saprophytic bacteria and yeasts of various
306
sizes and shapes, whilst smears taken from the deeper parts
generally do not show any bacteria.
The examination of the natives of the south coast of British New
Guinea revealed the presence of a great number of ulcers. Several
were well characterised and could be classified, whilst others did not
show any typical features.
TROPICAI, SLOUGHING PHAGEDAENA, ULCUS TROPICUM
Ulcus tropicum, a complaint occurring in practically every part
of the tropics, is known under nearly as many names as there are
localities where it has been found.
Clinically it is a chronic sloughing ulceration of phagedaenic
character, which may spread into the depth, laying bare muscles and
bones and showing a tendency to spontaneous healing, or leading,
if neglected, to considerable deformities of the affected extremities.
The literature on tropical sore is fairly extensive, and has been
summarised by Plehn (1914). It is, however, beyond doubt that
several varieties of sores, specific and non-specific, have been
previously included under the same heading.
Prowazek (1907) described a spirochaete, Spirochaeta schaudinni ,
as the specific cause of the disease. This spirochaete is actively
motile, resembling the balanitis spirochaete, possessing only rarely
periplast on one end; it multiplies by longitudinal fission and forms
involution forms.
The same spirochaete was found by Keysselitz and Mayer (1909)
in smears taken from the deep layers of ulcers, and by Wolbach and
Todd (1912); although the latter authors consider that 'The fact
that these ulcers are almost invariably found solitary and on regions
exposed to trauma supports the explanation that the process is a
reaction to an organism having slight power of invasion.’
Le Dantec (1911) had previously pointed out that two conditions
were essential for the development of a tropical sore, namely an
abrasion of the skin and exposure of this to soiling by humid earth.
A direct experimental transmission of the sore to animals such
as guinea-pigs, monkeys, and apes and to men has been
unsuccessful in the hands of all observers. Blanchard (1914)
succeeded in transmitting a tropical ulcer from man to man, but
307
only after having produced at first an artificial necrosis of the skin,
which was some days afterwards inoculated with the granulation
tissue from a case of tropical ulcer.
Cases of tropical ulcers were found in varying numbers,
throughout the coastal districts of British New Guinea, and barely
one village was entirely free from this complaint.
In the villages situated on high, dry ground only a small number
of cases was seen, whereas in the villages in the western parts of
New Guinea, which are built on swampy, muddy ground, a great
number of cases was encountered. It seems, therefore, that the
frequency of tropical ulcers stands in some relation to the incidence
of mangrove mud.
The tropical ulcer of New Guinea does not differ essentially from
that observed in other parts of the tropics. It occurs mostly on the
lower extremities, on the dorsal surface of the foot, in the neighbour¬
hood of the external and internal malleolus, on the skin, rarely on
the back of the thigh. In the early stages the ulcer resembles a boil.
There is an ulcer in the centre which is covered by a scab, slightly
raised above the surface of the skin, showing fluctuation and a
certain amount of inflammatory reaction around. Very soon, if
neglected, this small round ulcer begins to spread, the epithelial
layer breaks down, showing underneath a fungating ulcer, which
bleeds very readily and is covered with a small amount of thick
greyish pus. The ulcer is well defined from the surrounding tissues.
In the beginning the ulcer spreads very rapidly, is irregular in
outline, and covered by a layer of foul smelling greyish semi-fluid
pus, which forms a pseudo-membrane covering the dirty greyish to
reddish granulations (fig. 15). After a varying interval the ulcer
spreads into the depths, destroying the subcutaneous tissue and the
muscles (fig. 16) and often attacking the underlying bone, and gives
rise to periostitis and osteitis. Now and again a true osteomyelitis
may follow the ulcer, and then pieces of necrotic bone are often found
embedded in the granulation tissue. In other cases, however, the
ulcer remains superficial and the granulation tissue proliferates and
projects above the level of the surrounding skin. The borders in
chronic ulcers are either sharply defined or sometimes undermined.
The whole of the foot may swell up, resembling elephantiasis (see
Plate XXIII, fig. 17).
308
The ulcers are very chronic, and may persist for many years
without endangering the life or general health of the patients.
Judging by the number of natives seen with extensive scars above
the tibia and with thickening of the bone, it is evident that even
large ulcers of long standing may suddenly begin to heal up. At
other times the patients emaciate considerably, become completely
crippled, and consequently are confined to their houses and die of
the effects of septic resorption.
Treatment . Salvarsan has been introduced within the last few
years for the treatment of tropical ulcer with excellent results. This
mode of treatment is, however, too expensive and complicated to be
generally adopted. The thorough removal of the granulation tissue
with a sharp spoon, followed by careful dressing with a mixture of
iodoform-boracic acid in the proportion of I to io is efficient. The
majority of the ulcers yield readily to this treatment, and even large
sores may heal up within a few months.
Natives who have been in contact with Europeans for a longer
period bandage the sores with banana-leaves, and this treatment
seems to be fairly successful.
Pathological Histology of Ulcus tropicum
Opportunity was taken to obtain granulation tissue of io cases
suffering from Ulcus tropicum in various stages for further histo¬
logical examination. The tissue which was preserved in 80 per cent,
alcohol was embedded in paraffin and sections stained with
haematoxylin eosin, Breinl’s saffranin-methylene blue, orange-tannin
(Breinl, 1908), and by Heidenhain’s method, using iodine-potassium
iodide solution as a mordant.
The results of histological examination were, on the whole, in
accordance with those published by Keysselitz and Mayer and by
Wolbach and Todd.
The sections of the granulations from the centre of the lesions
consisted of a loose fibrous mesh-work, the meshes were filled with
exudate, containing red blood corpuscles, leucocytes and lympho¬
cytes, plasma-cells, and a few eosinophile leucocytes were scattered
here and there. The granulation tissue was richly vascularized,
many of the arteries showing endarteritis obliterans, and was covered
by a loose layer of fibrin, which contained erythrocytes in all stages
309
of disintegration, lymphocytes, cell detritus and bacilli and spiro-
chaetes in great numbers.
The sections from the border of the ulcer showed a marked
hypertrophy of the epithelial layer, being of many times its normal
thickness. In between the epithelial cells were roundish or oval
well-defined spaces of varying size, filled with a loose and
oedematous fibrous tissue, in which a small number of polymorpho¬
nuclear leucocytes were embedded. The epithelial strands between
these spaces became thinner, the cells showing degenerative changes,
increasing in size and becoming vacuolated. The nuclei became
irregular in shape and finally disappeared. In close proximity to
the ulcers these spaces were densely packed with leucocytes and
lymphocytes, resembling microscopically small abscesses.
The superficial epithelial cells from the edge of the sore showed
degeneration, the cytoplasm was vacuolated, and the nuclei were
irregular and did not stain evenly.
In the sections the same microorganisms as described by previous
observers were found. Fusiform bacilli occurred mostly on the
surface of the ulcer, and were found in abundance in between large
numbers of bacteria and cocci.
The spirochaetes —Spirochaeta schaudinni , Prowazek—to which
the etiology of ulcus tropicum is generally attributed, were seen in
small numbers in sections of several of the cases, mostly lying on the
surface between the other microorganisms or in between the epithelial
cells adjoining the ulcer.
CONTRACTING SORE
A special type of ulcer was encountered, cases of which were
irregularly distributed throughout the coastal districts of British
New Guinea. A comparatively greater number of cases was seen in
the eastern parts, but sporadic cases were discovered in practically
every district visited. This ulcer differed in its appearance consider¬
ably from other sores, and its clinical features were so well marked
and uniform that it was considered a hitherto undescribed form of
skin ulceration. It was named 1 Contracting Sore 1 on account of its
effects. It heals up with formation of dense scar tissue, leading to
contraction of the joints.
3io
The sore appeared in the majority of cases in the neighbourhood
of joints, for example on the inner or outer surface of the knee
(fig. 18), on the dorsal surface of the foot, above the tendo achillis,
or in the neighbourhood of the shoulder joints (fig. 19). Other
localities, such as around the anus (fig. 20), the abdomen or chest
or arms were only rarely the seat of the ulcer. On the whole it
occurred more frequently in young persons, and was often found in
children. Natives of more mature age only showed the typical scar
formations.
The earliest stage was in a boy about 10 years of age. A small
ulcer of about 3 mm. in diameter was found on the inner side of the
knee. It was round and possessed irregular burrowing edges; the
surface was covered by a dry looking, slightly raised, yellowish,
greasy scab; the surrounding tissue did not show any marked
inflammation. Underneath this scab, which could easily be
removed as a whole, was a collection of thick greyish pus accumu¬
lated above the granulating surface, which was slightly below the
level of the skin, of reddish colour, even and of the appearance of
coarse sandpaper. The granulation tissue was firm and could
easily be scraped off in strips by means of a sharp spoon.
The ulcers spread slowly and gradually, at first remaining
roundish, but after a varying period, becoming irregular in shape,
and were always covered by the same kind of yellowish-grey scab,
secreting from the edges an amber-coloured yellowish clear fluid.
They never spread below the underlying fascia, never attacked the
bone, and the granulation tissue could always be taken off in whole
strips, forming a layer of 2 to 5 mm. in depth and exhaling a
pungent, penetrating odour.
The ulcer may implicate in some natives a small area only and
the process may come to a standstill. More often extensive areas
may be affected, as in fig. 21, where the sore had spread over the
whole flexor surface of one leg.
At the time when the ulcerative process is still progressing there
may be in parts a marked tendency to spontaneous healing (see
Plate XXIV, fig. 22). The granulating surfaces clear up and are
replaced by dense and hard scar tissue, possessing a smooth surface,
and being often extensively intersected by slightly raised strands of
hard connective tissue. After a varying period, ranging from six
months to three to four years, the ulcer may completely heal up in the
way described, giving rise to contraction of the affected parts closely
resembling at first sight the scar formation due to deep burns (see
figs. 23, 24).
This contracture invariably causes further deformities (fig. 25),
such as complete ankylosis of the joints and wasting of the muscles
on account of inactivity.
Histology. From nine cases suffering from different stages of
1 Contracting Sore * granulation tissue was obtained by scraping the
sore with a sharp spoon. It was fixed in 80 per cent, alcohol and
sections made and stained by various methods. The granulation
tissue from an early ulcer about 3 cm. in diameter showed, on
microscopic sections, a marked hypertrophy of the epithelial layer
immediately surrounding the ulcer. These hypertrophied layers of
epithelial cells were interspersed with irregular spaces filled with a
loose oedematous connective tissue, the meshes of which contained
a small number of leucocytes and lymphocytes and a few plasma
cells; the nearer to the ulcer the larger were these spaces, the thinner
the epithelial strand separating them and the denser the cell
infiltration.
The corium was oedematous and contained a certain amount of
infiltration of lymphocytes, leucocytes and plasma cells, which
infiltration was denser around the blood vessels.
In the central parts of the ulcer the epithelial layer had been
replaced by granulation tissue, showing here and there a few
epithelial cells in varying stages of degeneration. The granulation
tissue itself did not show any distinctive features.
A number of large cocci and bacteria were seen in sections from
the surface of the ulcer lying between red blood corpuscles in different
stages of disintegration and coagulated fibrin. There were neither
fusiform bacilli nor spirochaetes in the surface layers. In the deepest
layer of the epithelium, however, in the rete mucosum of the
malpighian layer especially in the cells lining the spaces referred to,
a great number of spirochaetes were found, which were either intra¬
cellular or were lying in the fine clefts which separated the cells.
These spirochaetes occurred either singly or sometimes in bunches,
and stained readily with iron haemotoxylin after iodine-potassium
iodide had been used as a mordant. The spirochaetes were found
312
only in this epithelial layer, and were neither present in the granula¬
tion tissue nor in the discharge, of which a small number of smears
was examined.
The sections of the granulation tissue of advanced cases did not
show any especial features. It was richly vascularized, some of the
vessels showing endarteritis obliterans. There were numerous leuco¬
cytes and lymphocytes, a few plasma cells and many phagocytes
containing small roundish cell inclusions which stained blackish
with Heidenhain’s iron haematoxylin, and orange when Breinl’s
method was employed.
The deepest layers of the granulation tissue consisted of a mesh-
work of newly-formed dense connective tissue, with numerous fibro¬
blasts with a small amount of small-celled infiltration.
Judging from the histological examination, the ulcer begins with
a progressive destruction of the epithelial covering of the skin, whilst
the cells of the deepest layers of the rete mucosum are invaded by
numbers of spirochaetes. The deep layers of the granulation tissue
show a tendency to formation of a dense connective tissue.
On the whole the granulation tissue of these contracting sores
does not differ microscopically in any essential point from that of the
ulcus tropicum. The small-celled infiltration is much denser in the
latter, the surface layers contain a great number of microorganisms,
fusiform bacilli, bacteria and spirochaetes, whereas in the sections of
the contracting sore only a very small number of bacilli were found
in the surface layers, and the spirochaetes were only seen in and
between the epithelial cells of the rete malpighi.
The histological examination of the granulation tissue of the
contracting sore did not reveal any microorganism which could be
regarded as specific.
The presence of spirochaetes in numbers in and between the
layers of the epithelial cells of the malpighian layer is very
suggestive, but spirochaetes are found in great numbers in many and
various ulcers of the skin. Moreover, it was impossible to work out
any detailed morphology of the spirochaete with the material
collected.
3^3
ULCUS INTERDIGITALE DESTRUENS
A form of ulceration closely resembling ‘ Ulcus interdigitale * as
described by Castellani and Chalmers (1913) was seen among the
natives living in the swampy parts of the western New Guinea.
According to these authors, the ulcer begins as a fissure in
between the toes, which * Rapidly deepens and enlarges into a large
oval ulcer with a dull dark red fundus and sodden-looking margins.
There is practically no discharge whatsoever.’
The sores observed resembled, in some respects, those described.
The ulceration usually started as a fissure between the toes, or more
often in the sulcus below the toe (PI. XXV, fig. 26). This small fissure
formed, after a time, into a small painful ulcer, and spread fairly
rapidly upwards to the toe and downwards to the sole of the foot
(fig. 27). The ulcer is deep, possessing sharp irregular edges, and
the granulation tissue, which is covered by an irregular dirty greyish
scab, has a reddish uneven fundus and discharges a great deal of
thick yellowish pus. When the ulceration spreads upwards in
between the toes it causes, on healing, the two adjoining surfaces of
the toes to grow together. Sometimes the whole toe becomes
covered with granulation tissue which leads to complete loss of the
affected toe. Such an example is shown in fig. 28, where the
ulceration has spread to the sole of the foot, implicating the ball of
the foot, and had given rise to a deep, irregular granulating sore,
and two of the toes had been amputated by the ulceration.
These interdigital sores are very chronic, and show, after a
shorter or longer period, a tendency to spontaneous healing after
having caused considerable deformity of the foot. Fig. 29 was
taken from a case where the fourth and fifth toes had disappeared,
and where the second and third toes had grown together on their
adjoining surfaces.
Cases of this type of ulcer were widely distributed throughout
the districts of New Guinea west of Orokolo, and affected mostly
men. The ulcers differ from the ‘ Ulcus interdigitale’ on account of
their copious discharge and their tendency to lead to the destruction
of the affected toes, and the name ‘ Ulcus interdigitale destruens * is
proposed for them.
Pathological Histology. From two cases suffering from inter-
3*4
digital sores, granulation tissue was obtained by scraping the sore
with a sharp spoon.
Microscopically the sections did not show any characteristic
features. The epithelial surrounding of the ulcer was hypertrophic,
the granulation tissue which was richly vascularised showed a heavy
leucocytic and lymphocytic infiltration. Numerous bacteria and
large cocci occurred on the surface, lying in between fibrin cell
detritus and disintegrating red blood corpuscles.
GANGOSA
Gangosa or Rhinopharyngitis mutilans, a disease which causes
ulceration and destruction of the nose, soft and hard palate, and of
the skin of the face, is endemic in most of the coastal districts of
British New Guinea. A full account is being published in this
journal, describing Cryptococcus mutilans as its etiologic agent
(sec p. 213). In several far advanced cases the hands and feet were
the seat of extensive ulceration, the fingers were enlarged, the back
of the hands swollen and discharged copiously.
These lesions are in all probability due to the same etiologic
agent as the lesions of the face.
SKIN DISEASES
A great number of different skin diseases are prevalent amongst
the natives of New Guinea.
Tinea imbricata (Plate XXVI, fig. 30) was found in every district,
equally prevalent amongst those natives who had been civilised for
years, and amongst those who had not come in contact with
Europeans and had not been away from the district.
A few cases of Leucoderma were encountered, mostly confined to
the hands (fig. 31).
Only two cases of true Albinism were found in the Trobriand
Islands of the north-east coast of New Guinea—one was in an adult
man, the other in a baby about three years of age.
A peculiar form of skin disease was encountered somewhat
resembling Acne in its clinical appearance, and leading to destruction
and peculiar scar formation of the affected parts. Sporadic cases of
this complaint, occurring in men and women, were distributed
3*5
throughout British New Guinea. The nose, sometimes the upper
lip, both cheeks and the forehead, were covered with pustules from
which sebaceous material could be expressed. The nose became
enlarged, hyperaemic, and the skin uneven and covered with warty
excrescences. In one case the upper lip was similarly affected
(fig. 32). Here and there were small patches of smooth, shiny scar
tissue, and small roundish depressions with a reddish-looking
fundus, resembling to some extent the pitting after smallpox
(fig- 33 )-
This disease showed a tendency to destroy the affected parts.
The free border of the alae was uneven and irregular and, in two out
of eight cases, the soft palate and the uvula had been destroyed and
replaced by dense scar tissue.
The skin of the body of the patients was clean and did not show
any lesions whatsoever.
As far as histories could be obtained, the complaint seemed very
chronic, extending over years, and invariably healed spontaneously.
As far as could be ascertained, the disease made its first appearance
in the form of pustules on the alae of the nose, whence the lesions
spread gradually to the cheeks and upper lip.
An advanced active case is of typical appearance, but when the
lesions had partly healed the differential diagnosis now and again
offered difficulties.
The microscopic examination of the sebaceous material expressed
from pustules did not give any clue to the etiology.
CONCLUDING REMARKS
~~ The material collected during the journey has shown that a great
many diseases which occur commonly amongst the native popula¬
tions in other parts of the tropics are present also in New Guinea.
Of epidemic diseases only dysentery has made its appearance
within the last twenty years, and since 1896 outbreaks have been
recorded from different districts of the Possession.
A? the primary object of the expedition was the mapping out of
the distribution of diseases in New Guinea, the short stay did not
permit a thorough investigation into a number of complaints,
especially ulcers and skin diseases, many of which could not be
3*6
diagnosed and might, on further examination, prove new to
medicine.
The two expeditions were made possible by the practical interest
of the Department of External Affairs, Commonwealth of Australia.
I take the opportunity of expressing my indebtedness for courtesy
and assistance received from the Secretary of the Department of
External Affairs, Mr. Atlee Hunt, C.M.G., from the Lieutenant-
Governor of Papua, Judge J. H. P. Murray, C.M.G., and his
personal staff, and from the Government Secretary, Mr. A. M.
Campbell.
3*7
APPENDIX
LIST OF PAPUAN MOSQUITOS
*Anopheles (Myzorhynchus) barbirostris , var. bancrofti , Giles.
Anopheles ( Nyssorhynchus) annulipes , Walker.
Anopheles (Cellia) punctulata. Ddnitz-Theobald.
*Armigeres ob turbansf Walker.
Neosquamomyia breinli , Taylor.
* Stegomyia fas data, Fabr.
Stegomyia scutellaris , Walker.
Stegomyia pseudoscutellaris, Theobald.
Stegomyia omata , Taylor.
Stegomyia atra , Taylor.
Scutomyia notoscripta, Skuse.
Lepidotomyia lineatus , Taylor.
Leucomyia australiensis , var. papuensis, Taylor.
Leucomyia ? albitarsis. Taylor.
Culicelsa vigilax , Skuse.
Culex sitiens , var. mi Ini, Taylor.
Culex fatigans, Wied.
Pseudotaeniorhynchus conopas . var. giblini , Taylor.
Chrysoconops brevicellulus , Theobald.
Taeniorhynchus septempunctata , Theobald.
Taeniorhynchus uniformis, Theobald.
Taeniorhynchus papuensis, Taylor.
Melanoconion papuensis , Taylor.
Finlaya poicilia , Theobald.
Uranotaenia nigerrima . Taylor.
Hodgesia triangulatus , Taylor.
The bulk of the mosquitos, of which a list has been prepared by Mr. F. H
Taylor, were collected by Dr. Giblin and the writer at the Lakekamu gold field,
except those marked with an asterisk, which are previously recorded species.
(Taylor, 1914).
3*8
REFERENCES
Ashburn and Craig (1907). Observations upon Filaria pbilippensis and its development in
the Mosquito. Philippine Journal of Science , II, p. 1.
Bahr (1912). Filariasis and Elephantiasis in Fiji. London.
Blanchard (1914). Inoculationes experimentales de l’ulcire phagedenique tropical. Bull.
Soc. Path. Exot., VII, p. 96.
Breinl (1908). Method of preparing and staining wet films. Annals Trop. Med. and Paras.
I, p. 470.
Castellani and Chalmers (1913). Manual of Tropical Medicine. 2nd edition. London.
Le Dantec (1911). Precis de Pathologie exotique, Paris, 3rd edition.
Fontoynont et Carougeau (1918). Nodosites juxta-articulaires ; Mycose due au Discomyces
Carougeaui. Archives de Parasitologic , Vol. XIII, p. 583.
Fulleborn (1911). Untersuchungen iiber Mikrofilarien in der Siidsee. Archiv f. Scbiffs-
und T ropenbygiene , XV, p. 368.
-(1912). Ueber Mikrofilarien des Menschen im deutschen Sudseegebiete und deren
Turn us nebst Bemerkungen iiber die klinische Manifestation der dortigen Filariasis.
Arcbiv f. Scbiffs- und Tropenbygiene, XVI, p. 533.
- (1913). Beitrage zur Morphologic und Differential diagnose der Mikrofilarien. Arcbiv
f. Scbiffs- und T ropenbygiene, XVII, Beiheft 1.
Gros (1907). Nodosites juxta-articulaire de Jeanselme chez les indigenes musulman d’Algerie.
Arcbiv f. Scbiffs- und Tropenbygiene, XI, p. 552.
Jeanselme (1906). Des nodosites juxta-articulaires observees sur les indigenes de la presqu’ile
Indo-chinoise. Arcbiv f. Scbiffs- und T ropenbygiene, X, p. 5.
Joyeux (1913). Contribution A l’etude des nodosites juxta-articulaires. Bull. Soc. Path.
Exot., VI, p. 711.
Keysselitz und Mayer (1909). Ueber das Ulcus tropicum. Arch. f. Scbiffs- und Tropenbygiene,
XIII, p.137.
Ki nos hit a (1906). Ueber die Verbreitung der Anophelen auf Formosa und deren Beziehungen
zu den Malariakrankheiten. Arcbiv f. Scbiffs- und Tropenbygiene, X, p. 708.
Lebouef (1911). Note sur l’existence des nodosites juxta-articulaires de Jeanselme dans
l’Archipel caledonien. Ann. Hyg. et Mid. Col., p. 549.
Lynch (1905). Brit. Med. Jour., p. 21.
MacGregor (1897-1898). Annual Report of Papua, Sanitary, p. XXXIV.
Neveux (1907). Le Narinde, fibromatose sous-cutanee des Toucouleurs du Boundou (Senegal).
Rivue Mid. et Hyg. Trop. IV, p. 183.
Plehn (1914). Die tropischen Hautkranlcheiten. Handbucb der Tropenkrankbeiten, Leipzig,
Vol. II, p. 171.
Prowazek (1907). Vergleichende Spirochaetenuntersuchungen. Arb. a. d. Kaiserl. Gesund -
beitsamt, XXVI, p. 23.
Steiner (1904). Ueber multiple subkutane harte fibrose Geschwiilste bei den Malayen.
Arcbiv f. Scbiffs- und Tropenbygiene, VIII, p. 156.
Taylor, F. H. (1914). Culicidae from Papua. Trans. Ent. Soc. London, Part I, pp. 185-205.
Pis. XVII, XVIII.
Taylor, F. H. (1914). Contributions to a knowledge of Australian Culicidae, No. 1. Proc.
Linn. Soc. N. S. Wales, Part 3, pp. 454-468. Pis. XXXIV, XXXVII.
Thorpe (1896). Brit. Med. Journ., II, p. 922.
Wolbach and Todd (1912). A study of chronic ulcers, Ulcus tropicum, from the Gambia.
Journ. Med. Research, XXII, p. 27.
320
EXPLANATION OF PLATES
Plate XIX
Fig.
i.
Elephantiasis of scrotum.
Fig.
2.
Elephantiasis of vulva.
Fig.
3-
Leprosy; advanced case.
Fig.
4-
Juxta-articular nodules.
Annals Trop. Med. o Parasitol ., Vol. IX
PLATE XIX
Fig. i
Fig. 3
F 'c- 2 Fig. 4
C. Tinting Co., Ltd., Imp.
322
Plate XX
Fig 5- Juxta-articular nodules.
Fig. 6. Yaws. General eruption. Secondary stage.
Fig. 7. Yaws. General eruption. The same case as fig. 6.
Fig. 8. Yaws. Note destruction of nose.
Fig. 5 Fig. 6
Fig. 7 Fig. 8
C. Tin ling &* Co., Ltd.., Imp.
3 2 4
Plate XXI
Figs. 9-12. Disease characterised by osteitis, periostitis, arthritis,
and formation of sores.
Fig. 9. Late stage, resembling Madura foot.
Fig. 10. Swelling of bone affecting the tibia.
Fig. II. Swelling of bone affecting the diaphysis of radius.
Fig. 12. Mutilation of hand after resorption of metacarpal
bones.
Annals Trop. Med. & Parasitol ., Vol. IX
PLATE XXI
Fig. ii
Fig. 12
C. Tinling Co., Ltd., Imp.
326
Plate XXII
Figs. 13-14. Disease characterised by osteitis, periostitis, arthritis,
and formation of sores.
Fig. 13. Metatarsus of large toe disappeared, the toe in
hyper-extension and abduction.
Fig. 14. Late stage of disease, showing irregular scar
formation.
Figs. 15-16. Ulcus tropicum.
Annals Prop. Med. & ParasitolVol. IX
PLATE XXII
Fig. 14
C. Tinling Co., Ltd., Imp .
PAGE NOT
AVAILABLE
Plate XXIII
Fig. 17. Ulcus tropicum.
Figs. 18-21. Contracting sore.
Annals Trop. Med. G? Parasitol., Vol. IX
PLATE XXIII
Fie. 19
Fig. 21
Fig. 20
Fig. 17
Fig. 18
C. Tinling Co., Ltd., Imp.
33 °
Figs. 22-25.
Plate XXIV
Contracting sore.
Annals Trop. Med. £*? ParasitolVol. IX
PLATE XXIT
%
x
•v
Fig. 24
Fig. is,
C. Tift ling <5^ Co.. Lid., Imp.
332
Plate XXV
Figs. 26-29. Ulcus interdigitale destruens.
Annuls 'Trot>. Med. ParasitolVol. I .Y
PLATE XXV
Fig. 27 Fie. 29
C. Tin ling «$-* Co., Ltd., Imp
-
PAGE NOT
AVAILABLE
334
Plate XXVI
Fig. 30. Tinea imbricata.
Fig. 31. Leucoderma.
Figs. 32-33. Skin disease resembling Acne.
Annals Tro[>. Med. ZA Paras itol., Vol. IX
PLATE XXVI
Fic. 31 Fig. 33
C. Tinting CoLtd., Imp.
335
INSECT FLAGELLATES AND THE
EVOLUTION OF DISEASE, WITH
REMARKS ON THE IMPORTANCE
OF COMPARATIVE METHODS IN
THE STUDY OF PROTOZOOLOGY
BY
H. B. FANTHAM, M.A. Cantab., D.Sc. Lend.
LECTURER ON PARASITOLOGY, LIVERPOOL SCHOOL OF TROPICAL MEDICINE
{Received tor publication l May , 1915)
CONTENTS
PAGE
I —Introduction . 335
II —The Significance of the Herpetomonad Stage of Ltisbmania . 337
III — The Existence of the Herpetomonad Stage of Leisbmania and Allied
Parasites in Man . 339
IV — The Presence of Herpetomonads in Nature in other Vertebrates ... 339
V— The Occurrence of Herpetomonads in Plants . 341
VI —Experiments with Insect Flagellates belonging to the Genera Hcrpetomonai
and Critbidia . 342
VII — Some Inferences . 344
VIII — Summary . 346
References . 347
I. INTRODUCTION
The parasitic theory of disease has been greatly extended and
consolidated during the last generation. In the domain of
bacteriology it has perhaps become best known, at any rate to the
general public. Vast strides have been made during the latter half
of the period in our knowledge of animal parasites, especially of the
inter-relationship of Protozoa and arthropods in the propagation of
disease. There is also a re-awakening as to the importance of
mycology, or the study of fungoid organisms as pathogenic agents
of economic importance.
In the domain of protozoology, with which the present paper is
concerned, the progress, although rapid, has not been so great as it
might have been. Some of the reasons for this are that the work
is in the main in the hands of medical men whose knowledge of
zoology is generally of the slightest and their acquaintance with the
comparative aspect of the subject just as limited, while the few
zoologists engaged have in some cases drifted into a narrow outlook
and become lost in academic terminologies and discussions. The
result is that though many important discoveries have been made,
they have not reached full fruition, chiefly on account of lack of
application of the comparative method. As an example, the parasite
of kala-azar may be considered. The causal organism, Leishmania
donovani , was described in 1903 by Leishman and by Donovan,
while Rogers in 1904 cultivated it and discovered the herpetomonad
stage. The latter discovery, that of Rogers, was of fundamental
importance, but its application is, even to-day, only realised in
varying degrees by two or three British and French workers.
However, the output of literature on kala-azar and other
leishmaniases is simply enormous, though the progress resulting
therefrom is very slight, because of the overburdening amount of
controversy obscuring the issue. When Rogers in 1905 suggested
the bed-bug as the probable transmitter of kala-azar in India, and
Patton began to work at the same, the results obtained were largely
buried in criticism, much of it speculative and irrelevant. We read
in 1912 such statements as the following : ‘ It is quite clear that the
gut of the bug when it contains blood acts as a culture-tube
in which various organisms can live and develop. . . . Hence the
fact that the leishmania of kala-azar and oriental sore become
flagellates in the bug’s gut is in itself no proof whatever that
these insects are the transmitter’s of these diseases.* It is to be
regretted that those putting forward or supporting the latter
remarks did not more carefully consider the suggestions and
inferences to be drawn from cultural methods, and proceed forthwith
to experiment directly with the herpetomonads found in insects,
especially after Patton (1907) had given the first account of the
complete life-cycle of such a herpetomonad, and Darling (1910) had
suggested that a case of oriental sore in Panama had resulted from
' an inoculation with an invertebrate gut flagellate (Crithidia ?).*
Again, we had the flood-gates of destructive criticism opened when
Basile (1910-11) suggested that Leishmania infantum was
transmitted by fleas, though it does not seem to have occurred to
337
any of the critics to begin experiments on the effects of flagellates of
fleas when introduced into the vertebrates they infest. However, it
is unnecessary to labour the point further, it is merely symptomatic
of the age, pointing to the partial failure of the modern educational
system, in that it tends to the development of trifling academic
debate, nearly always useless and sometimes even harmful, and to a
fostering of the partisan spirit, wherein a broad comparative outlook
is seldom attained. Legitimate criticism is always welcome, but
when it degenerates into mere retort and negation it is subversive
of progress.
There is yet another branch of the study of pathogenic organisms
which has not yet received attention, namely, the biological evolution
of such forms as disease producers, and a consideration of ‘ disease
in the making.* For this study a knowledge of comparative
morphology and of the inferences to be made therefrom is absolutely
essential, and any one who aspires to become a parasitologist must
be prepared to undertake such a study.
II. THE SIGNIFICANCE OF THE HERPETOMONAD STAGE OF
LEISHMANIA
When Rogers in 1904-05 cultivated the Leishman-Donovan body,
obtaining a herpetomonad flagellate therefrom, and suggested that
the genus Leishmania should give place to Herpetomonas , a great
step forward might and should have been made in the application
of the idea, especially as Christophers (1904) in L. donovani , and
Mesnil and colleagues (1904) in L. tropica , had shown that a
structure, now known to be the root of the flagellum, could be found
in some of the parasites. ** Patton (1908) also placed Leishmania
donovani in the genus Herpetomonas. It was also pointed
out that the natural herpetomonads of insects might be mistaken
for developing Leishmania. However, these advances met
with the inevitable destructive criticism, for as recently as 1912
we read: ‘ The genus Herpetomonas includes flagellates which
have only one, and that an invertebrate host. The fact that
the parasite of kala-azar has two hosts and can live and
multiply in the organs of a vertebrate shows it to be profoundly
• Novy (1909) »howed that a rhizoplast occurred also in /.. infantum. (Bull. Soc. Path.
Exol., II, pp. 385-387.)
338
different from the true Herpeiomonas , which lives only in the
invertebrate. This fact alone would be sufficient to establish the
genus Leishmania .When a Herpetomonas becomes so
changed that it has acquired the power of living and developing as
an intracellular parasite in the body of a warm-blooded animal,
and giving rise to such a serious disease as kala-azar,
we are justified in concluding that it has passed out of the
genus Herpetomonas , from which it originated, into the genus
Letshmania* The pity is that the author, before penning
these remarks, did not himself endeavour to determine experi¬
mentally whether Herpetomonas might not acquire the power
of living and developing in a vertebrate, and that the record of the
remarkable observations of Dutton and Todd, published in 1903,
on the presence of herpetomonads in the blood of Gambian house
mice was unknown to him, as to many others. I deeply regret the
necessity for making these remarks; there is nothing personal in
them, but the facts are there and cannot be gainsaid.
The successful cultivation of L. tropica and L. infantum again
did not advance matters, but discussion arose as to the number of
valid species of Leishmania y and numerous attempts were made to
inoculate the viruses into laboratory mammals. Although expedi¬
tions were organised for the study of Leishmania in endemic areas,
it never seems to have occurred to those working thereon to try
to introduce herpetomonad flagellates of insects into vertebrates.
Apparently, vertebrates, being thought much more powerful and
important animals than invertebrates, must perforce be primarily
considered and experimentation undertaken with them viewed
as the principal hosts. Unfortunately, human suffering has paid
dearly for this one-sided view, which is only equalled by the
ridiculous arguments, that used to occupy the attention of amateur
parasitologists and waste the time of learned societies, as to which
was the primary or principal host of a digenetic parasite, was it the
vertebrate or the invertebrate host ? The answer is that each host
has its own particular importance which must be carefully considered.
In the case of the kala-azar parasite the pathogenic organism
certainly seems to have arisen from a herpetomonad parasite of an
invertebrate (probably an insect), and the flagellate has acquired the
capacity to live within a vertebrate and so become digenetic.
339
Again, the intimate connection of natural canine leishmaniasis
and of infantile kala-azar has been explained away by some. In
other words, every step forward has been obscured by irrelevant
criticism, and what should have been obvious inferences were either
ignored or never given the opportunity of being put to the practical
test. The comparative aspect was overlooked. The occurrence of
natural herpetomonads in invertebrates must not only be acknow¬
ledged, but it must be allowed that they may become pathogenic
when introduced into vertebrates.
III. THE EXISTENCE OF THE HERPETOMONAD STAGE OF
LEISHMANIA AND ALLIED PARASITES IN MAN
It is now known that Escomel in Peru in 1911 found the herpeto-
monad stage of Leishmania tropica in man, but seeming somewhat
disturbed by his finding did not publish it at the time, nor realise
its importance and significance. Monge, also in Peru, records
(1914) the presence of the herpetomonad stage in dermo-mucosal
leishmaniasis, and La Cava (1912) in L. tropica in Italy. Splendore
(1912) saw elongating forms and a few flagellate stages in cases in
Brazil.
Even before these observations, Darling (1906, 1909) in Panama
had recorded a disease resembling kala-azar, and described the
causal parasite therein as His toplasma capsulatum. He stated that
flagellate stages occurred in the lungs and spleen. As might be
expected, the mention of the occurrence of flagellate stages passed
almost unnoticed. In 1912 Rocha-Lima, working in Hamburg,
stated that in his opinion Histoplasma capsulatum was a
Blastomycete. Whether this be so or not, surprisingly little
attention was paid to Darling’s researches.
Again, in 1913 Franchini described a herpetomonad parasite
from man, and called it Haemocystozoon brasiliertse.
IV. THE PRESENCE OF HERPETOMONADS IN NATURE IN
OTHER VERTEBRATES
Dutton and Todd in 1903 recorded the presence of herpetomonad
flagellates in the blood of mice in the Gambia. Unfortunately, these
observations, first made in 1902, seem to have escaped general
notice. They have since been interpreted otherwise by the surviving
34 °
author. Three Gambian house mice out of fourteen were found to
be infected with flagellate herpetomonads, of which a clear description
is given, together with dimensions. The infection was scanty, and
the parasites were only seen in fresh preparations. The organisms
were definitely compared with Herpetomonas ( Leptomonas)
biitschlii recorded from the Nematode worm, Trilobus gracilis .
A diversion is necessary here. The parasite, Herpetomonas
( Leptomonas) biitschlii , briefly described by Biitschli in 1878 and
named by Saville Kent in 1880, emphatically needs re-investigation.
According to Biitschli, the organism was spindle-shaped or almost
rod-shaped, bearing a long flagellum and possessing a contractile
vacuole—a feature not present in flagellates latterly known as
‘ leptomonads.* No nucleus was observed by him. A darker,
refractile granular mass behind the contractile vacuole may have been
the blepharoplast, or even the nucleus. Saville Kent appears to have
considered it to be the nucleus, and he incorrectly reproduced it as a
vesicle in his ‘ Manual of the Infusoria.* The developmental cycle
was unknown to Biitschli, and still remains so. The organism has
not been studied by modern methods of technique, probably because
of the difficulty of obtaining the host. It seems rather remarkable
that a parasite which is somewhat shrouded in mystery should be
made the type species of a genus, Leptomonas , which some workers
would like to see supersede the well-known genus Herpetomonas .
Moreover, Biitschli in 1884, in his account of the Mastigophora in
Bronn*s ‘Tierreich,* considered the two generic names to be
synonymous and retained the name Herpetomonas. The account of
H. muscae domesticae by Prowazek having been shown by many
competent investigators to be inaccurate—the so-called bi-flagellate
characteristic being due to division—there is no need for the
multiplication of genera and controversy on the same.
To return to the natural occurrence of herpetomonads in verte¬
brates. That these flagellates may occur naturally in mice is
undoubted, as they have been seen and described by other workers
in addition to Dutton and Todd. In May, 1909, while working in
the Quick Laboratory, Cambridge, a Herpetomonas was found in
life in the fresh peripheral blood of a mouse. It was seen almost
simultaneously and independently by Prof. Nuttall, Dr. Porter and
myself, but the infection was slight. The parasite has been seen
since, but again the infection was scanty. These observations have
just been published by Fantham and Porter. The parasite is very
probably Herpetomonas pattoni (Swingle), first seen in rat-fleas, as
some of these ectoparasites were breeding in the rat cages in the
animal house in which the mice were kept.
The so-called herpetomonad stages of trypanosomes are probably
natural herpetomonads of vertebrates co-existing with trypanosome
infections. This subject is discussed on page 345.
V. THE OCCURRENCE OF HERPETOMONADS IN PLANTS
In 1909 Lafont announced the discovery in Mauritius of a
Herpetomonas —or Leptomonas , as he called it—in the latex of a
species of Euphorbia or spurge. Confirmation followed rapidly, and
the flagellate (H . davidi) is now known in various species of
Euphorbia in neighbouring islands, in India, Africa, Portugal, and
other places (see Franca, 1914). In some of these places Hemiptera
were found crawling over the plants, and herpetomonads were also
found in the gut of the Hemiptera. Some of the parasitised plants
were diseased, and the malady has been termed 4 flagellosis.*
The flagellate parasite of the spurge plants is probably an insect
flagellate, coming from the Hemiptera infesting the plants. The
herpetomonad has probably secondarily invaded the plant tissue,
having adapted itself so that it is now able to live in the latex of the
plants. Encysted or post-flagellate stages of H. davidi have been
obtained in cultures.
Nearly three years ago I was informed, by a competent
authority, that a number of Euphorbia containing herpetomonads
grew outside a certain hospital situated in an area in which kala-azar
was endemic, and in which kala-azar patients were being treated.
The shrubs were infested by insects. It seems remarkable that no
attempt was made to trace a possible connection between the plant
herpetomonad and kala-azar, doubtless such a possibility was
considered too remote. Remarks of mine regarding a possible
connection were 'received by my informant with polite incredulity,
which is not surprising, since the wisdom of lecturing on
Herpetomonas and Crithidia to students of tropical medicine has
been questioned more than once.
34 2
VI. EXPERIMENTS WITH INSECT FLAGELLATES BELONGING
TO THE GENERA HERPETOMONAS AND CRITHID1A
Thanks to the remarkable series of experiments recently carried
out by Laveran and Franchini in Paris, and by Fantham and Porter
in England, we now know that a number of species of Herpetomonas
and Crithidia , naturally occurring in insects, may be successfully
inoculated into or fed to mammals, especially rats and mice. The
flagellates live and multiply in the vertebrates and become pathogenic
thereto, the symptoms chiefly resembling those of kala-azar. It
must not be supposed, however, that success is attained in every
experiment, though a large proportion are positive, especially if the
vertebrate hosts are young. (Compare the prevalence of leishmaniasis
in young people (children) in the Mediterranean region.)
In the insect host the Herpetomonas or Crithidia is a natural or
specific parasite, and its effects on its host are not marked. The
life-cycle of the organism in the invertebrate consists of a
leishmaniform pre-flagellate stage, gradually growing into a
flagellate form and followed by a resistant post-flagellate,
leishmaniform stage adapted for extra-corporeal life and for
transmission to a new host. When such a parasite finds its way,
either by inoculation or by feeding, into a susceptible vertebrate, it
can assume again leishmaniform or flagellate facies. The mode of
infection of the vertebrate in nature seems to be contaminative, either
by its food or through an already existing abrasion or puncture on
the surface of its body. Cases in which the flagellate-infected
insects have been allowed to suck the blood of vertebrates have proved
negative. Further, Fantham and Porter have brought forward
experimental evidence which shows that post-flagellate forms of the
parasites are best adapted to begin life in a vertebrate host.
In certain cases leishmaniases may be derived from reservoir
animals, such as man and various other warm- and cold-blooded
vertebrates (see page 344), in other cases they may arise de novo.
Laveran and Franchini have successfully introduced into
mammals Herpetomonas ctcnocephali from the gut of the dog-flea,
H . pat tout from the gut of the rat-flea, Crithidia jasciculata from the
gut of Anopheles maculipennis, and C. melophagia from the gut of
the sheep-ked.
Experiments have also been made with H. muscae domesticae.
The mammals used were chiefly rats and mice.
3+3
Fantham and Porter (1914) went further, and showed that
flagellates occurring in insects unassociated with the experimental
vertebrate may be introduced therein. Their conclusions were as
follows: —
1 1. Insect flagellates, e.g., Hcrpetomonas jacutum (Leger)
from Nefa einerea , and Herpeiomonas ctenocephali
(Fantham), parasitic in the dog-flea, Ctenocephalus cants ,
can live inside certain vertebrates (e.g., mouse and dog,
respectively) and can multiply therein. This we have
shown experimentally.
‘ 2. If such flagellates be inoculated intraperitoneally, or are
fed by the mouth in food, the flagellates can find their way
into the blood stream and internal organs (e.g., liver,
spleen, bone-marrow) of the vertebrate host.
‘ 3. The insect flagellates are pathogenic to the vertebrates
experimented upon, producing symptoms like those of
leishmaniasis (kala-azar).
‘ 4. The oval post-flagellate forms appear to be more capable
of developing in vertebrate hosts than are other stages of
tjie herpetomonad parasite of the insect.
‘5. It may be expected that the various leishmaniases,
occurring in different parts of the world, will prove to
be insect-borne herpetomoniases.* (According to some,
leptomoniases would probably be considered correct, but
see page 340.)
In a later paper further experiments were recorded by Fantham
and Porter, and the researches were also extended to cold-blooded
vertebrates. The conclusions were : —
* 1. Herpetomoniasis can be induced in various warm- and cold¬
blooded vertebrates when the latter are inoculated or fed
with herpetomonads occurring in the digestive tracts of
various insects. The infection produced and the protozoal
parasites found in the vertebrates resemble those of human
and canine leishmaniases.
* 2. An infection can also be induced in certain vertebrates
when they are fed or inoculated with Crithidia gtrridis ,
and both flagellate and non-flagellate stages occur therein,
but no transition to a trypanosome was found.
3+4
‘ 3. The following Flagellata have proved pathogenic to warm¬
blooded mammals when the latter have been fed, or
inoculated subcutaneously or intraperiloneally with them—
Herpetomonas'jaculum , H. stratiomyiae , H. pediculi and
Crithidia gerridis . The hosts used were mice of various
ages. That H. ctcnoccphali can infect dogs has already
been shown by us.
‘4. Herpetomonas jaculum and Crithidia gerridis have also
been successfully fed or inoculated into cold-blooded
hosts, namely, fishes ( Gasterosteus aculeatus ), frogs,
toads, lizards ( Lacerta vivipard) and grass snakes
(Tropidonotus natrix).
‘5. As we have previously stated, we believe that leishmaniases
are arthropod-borne herpetomoniases, and that these
maladies have been evolved from flagellates of inverte¬
brates (especially herpetomonads of insects) which have
been able to adapt themselves to life in vertebrates.
4 6. In areas where leishmaniases are endemic, an examination
should be made of all insects and other invertebrates likely
to come into contact with men or dogs or rats and mice,
in order to ascertain if these invertebrates harbour herpeto¬
monads. Preventive measures should be directed against
such invertebrates, especially arthropods. Further, it is
likely that certain vertebrates, such as reptiles and
amphibia (especially those that are insectivorous), may
serve as reservoirs for leishmaniases or, as they should
preferably be termed, herpetomoniases. From such
reservoirs the herpetomonads may reach man by the
agency of ectoparasites or flies, especially such as are
sanguivorous.*
VII. SOME INFERENCES
The immediate and important inference to be drawn from the
experiments recorded in the previous section is that in them we sec
4 leishmaniasis in the making,’ as was pointed out by Fantham
and Porter in December, 1913, in these Annals. Also canine
leishmaniasis is probably herpetomoniasis due to H. ctenocefhal'i
in dogs.
A further imperative inference is that probably only one species
3+5
of Herpetomonas is concerned in adapting itself to life in vertebrates
in different parts of the world. This species is known under various
names, such as H . pattoni , H . ctenocephali , H. pediculi , and also as
H . donovani , //. infantum , //. tropica . These are probably merely
physiological races of a herpetomonad which is very like //. jaculum ,
briefly described by L6ger in 1902 from the gut of the Hemipteran,
cinerea. This herpetomonad under different conditions of
environment produces pathogenic effects in very varying degrees in
different vertebrates, from zero as in Dutton and Todd’s mice to
high mortality as in Indian kala-azar, and probably zero again in
cold-blooded hosts. It is also a flagellate which can probably live
in invertebrates not already recorded as being infected.
At various times statements have been made that herpetomonad
stages of trypanosomes occur in cultures. The invertebrate life-
cycles of all the trypanosomes, which so far have been well investi¬
gated, lend no support to the presence of a herpetomonad stage
therein. It is highly probable that the so-called herpetomonad
stages of trypanosomes were really cultures of scanty herpetomonad
infections co-existing with trypanosome infections. -It is known
that such invertebrates as rat-fleas may be infected with both
herpetomonads and trypanosomes at the same time. The different
discussions in the past, often acrimonious, as to what form of
flagellate occurring in an invertebrate constituted part of the life¬
cycle of a trypanosome or otherwise, are thus to be deplored.
Before concluding, attention may be drawn to some remarkable
and far-seeing suggestions regarding kala-azar published by
Rogers in 1905. He wrote that 4 the stomach of some blood¬
sucking insect is the most likely place in which to find
the natural development of the extra-corporeal stage of the
parasite, and that some such insect is the most likely carrier
of the infection.But as to the most likely kind of insects
to carry the infection we are on more uncertain ground, for
experiment alone can determine this. Nevertheless, it is worth while
to discuss which are the most probable kinds in order that
precautions may be taken against them without waiting for absolute
proof to be obtained.’ Especial notice is drawn to the last sentence,
published ten years ago, but not yet acted upon fully from the point
of view of preventive measures. However, exception must be made
for the excellent researches and preventive measures undertaken by
346
Dodds Price, at the suggestion of and latterly in collaboration with
Rogers, in the Assam tea gardens. He has reduced the mortality
due to kala-azar enormously by segregating the infected, by moving
coolie lines about 300 yards from older infected ones, and by having
new coolie lines placed on clean sites.
It has remained for workers in laboratories in temperate zones,
away from leishmaniasis material, again to point the way to
preventive measures and to indicate the origin of the disease.
VIII. SUMMARY
The significance of the herpetomonad stage of Leishmania y of
the recent announcements that such stages occur in man, and of the
presence of natural herpetomonads in other vertebrates (for example,
mice), are discussed. It also recalled that insect herpetomonads can
invade and live in plant-tissues.
The experiments on the introduction into different vertebrates of
various species of Herpetomonas and Crithidia parasitic in insects
by Laveran and Franchini, using mammals, and by Fantham
and Porter, using both warm- and cold-blooded vertebrates, are
summarised and discussed.
It is inferred that the various leishmaniases are due to a
herpetomonad of invertebrates which, under different conditions of
environment, produces pathogenic effects in very varying degrees in
different vertebrates, from zero, as in the mice described by Dutton
and Todd in 1903, to high mortality as in Indian kala-azar, and
probably zero again in cold-blooded hosts. It is also a flagellate
which can probably live in invertebrates not already recorded as
being infected. A human reservoir of leishmaniasis may occur in
some places, while warm- and cold-blooded vertebrates may also
function as the same.
It is highly probable that the so-called cultural herpetomonad
stages of trypanosomes were really cultures of scanty herpetomonad
infections co-existing with trypanosome infections.
It is recalled that Rogers in 1905 published that ‘it is worth
while to discuss which are the most probable kinds [of insect
transmitters of kala-azar] in order that precautions may be taken
against them without waiting for absolute proof to be obtained.'
Although these remarks were published ten years ago, little has been
done directly in the way of the preventive measures suggested. A
notable exception, however, is the work of Dodds Price in Assam.
347
Addendum. —June 28, 1915.
Since writing this paper my attention has been drawn to a recent
note by Archibald, who writes; ‘ Epidemiological and experimental
evidence does not support the theory that kala-azar in the Sudan is
transmitted by a biting insect. A more probable source of infection
appears to be some intermediate host, whose habitat is in water/
{Kept, of Advisory Committee for Trop. Dis. Resch. Fund for 1914,
p. 116 —published in April, 1915.) In this connection, see my
remarks on the contaminative method of infection in paragraph 2
on page 342.
REFERENCES
Further reference* will be found at the ends of some of the memoirs cited.
Basils, C. (1910). Sulla Leishmaniosi del cane e sulP ospite intermedio del Kala-Azar infantile.
Rendiconti R. Accad. dei Lincei , (ser. V.), XIX, ii, pp. 523-527
-(1911)* Sulla Leishmaniosi e sul suo modo di trasmissione. Rendiconti R. Accad. dei
Lincei , (ser. V.), XX, i, pp. 278, 479, 955. Sec also p. 50.
Botschli, O. (1878). Beitrage zur Kenntniss der Flagellaten und einiger verwandten
Organismen. I. Zeitscbr. f. toissen. Zoologie, XXX, pp. 205-281. 5 plates.
Christophers, S. R. (1904). On a parasite found in persons suffering from Enlargement
of the Spleen in India—second report. Sci. Mem. Govt. India , No. 11.
Darling, S. T. (1909). The Morphology of the Parasite ( Histoplasma capsulatum) and the
Lesions of Histoplasmosis, a fatal disease of Tropical America. Journ. Exptl. Med.,
XI, pp. 5 * 5 - 53 *- splato.
*- (1910). Autochthonous Oriental sore in Panama. Trans. Soc. Trop. Med.
® Hyg > rv, pp. 60-63.
Dutton, J. E., and Todd, J. L. (1903). ‘ Flagcllata in the Blood of a Mouse,' being part of
Sect. VII of the First Report of the Trypanosomiasis Expedition to Senegambia (i 9 ° 2 )-
Liverpool Scb. Trop. Med., Memoir XI, pp. 56-57.
Escomel, E. (1914). Leishmania flagelada en el Peru. La Cronica Medica. [Lima], XXXI,
pp. 224-227. See also (1913) Bull. Soc. Path. Exot ., VI, p. 237.
Fantham, H. B. (1912). Some Insect Flagellates and the Problem of the Transmission of
Leishmania. Brit. Med. Journ., Nov. 2, 1912, pp. 1196-1197.
Fantham, H. B., and Porter, A. (1913). Herpetomonas stratiomyiae , n.sp., a flagellate
parasite of the flies, Stratiomyia chameleon and 5 . potamida, with remarks on the biology of
the hosts. Annals Trop. Med. and Parasitol., VII, pp. 609-620. 1 plate.
-(1914-15). Some Insect Flagellates introduced into Vertebrates. Proc.Camb.
Pbilosopb. Soc., XVIII, pp. 39-50. 1 plate. Also Cambridge Univ. Reporter , XI.V,
p. 323 (Dec. 1, 1914).
-(1915). Further Experimental Researches on Insect Flagellates introduced into
Vertebrates. Proc. Camb. Pbilosopb. Soc., XVIII, pp. 137-148. Also Cambridge Univ.
Reporter , XLV, p. 931 (May, 25, 1915).
-(1915). On the Natural occurrence of Herpetomonads (Leptomonads) in Mice.
Parasitology, VIII, pp. 128-132.
3+8
Franca, C. (1914) Fa Flagellose des Euphorbes. Arch. f. Protistenkunde , XXXIV,
pp. 108-132. 1 plate.
Francium, (». (1913;. L’n nouveau protozoairc parasite de l’homme provenant du Brasil.
Bull. Soc. Path. Fxot VI, pp. 156-158. See alio pp. 333-336.
La Cava, F. (1912). De la Leishmaniose dci muqueuses et de la premiere dFcouverte de la
Lcisbmania tropica flagellcc dans lc corps humain. Bull. Soc. Path. Exot ., V, pp. 808-812.
Lav Fran, A., and Franchini, G. (1914). Infections de mammiferes par des Aagcllts d’invcrtF-
bres. Bull. Soc. Path. Exot., VII, pp. 605-612.
Mesmi., F., Nicolle, M., and Remlinger, P. (1904). Sur lc protozoaire du bouton d’Alep.
C.R. Soc. Biol., LVII, pp. 167-169.
Mover., l\ (1914). La Leishmaniasis del dermis en el Peru. Espundia, Uta, Juccuya, Qcepo,
Tiacc-arana. La Cronica Medica. [Lima], XXXI, pp. 231, 251, 288, 385.
Patton, W. S. (1907;. The Development of the Leishman-Donovan parasite in Cimex
rotundat us. Second report. Set. Mem. Govt. India, No. 31. Sec also No. 53.
-(11)07). Preliminary Note on the Life-cycle of a Species of Herpetomonas found in
Culex pipiens. Brit. Med. Journ ., July 13, 1907, pp. 78-80.
— (1908). Inoculation of dogs with the parasite of Kala azar ( Herpetomonas [Leisbmania]
donmant), with some remarks on the genus Herpetomonas. Parasitology, I, pp. 311-313.
Porter, A. (1914). The Morphology and Biology of Herpetomonas patellae, n.sp., parasitic
in the Limpet, Patella vulgata , together with remarks on the pathogenic significance
of certain Flagellates found in Invertebrates. Parasitology, VII, pp. 322-329.
Price, J. Dodds, and Rogers, Leonard (1914). The uniform success of segregation measures
in eradicating Kala-azar from Assam tea gardens. Its bearing on the probable mode
of infection. Brit. Med. Journ., Feb. 7, 1914, pp. 285-289.
Rocha-Lima, H. da (1912). Histoplasmosis und epizootische Lymphangitis. Arch. j.
Scbiffs- it. Tropenbvgiene. XVI, Bcibeft 1, pp. 79-85.
Rogers, L. (1905). The Conditions affecting the Development of Flagellated Organisms
from Leishman bodies and their bearing on the probable mode of Infection. Lancet ,
June 3, 1905, pp. 1484-1487.
Spi.endore, A. (1912). Lcishmaniosi con localizzazione nelle cavita mucose (nuova forma
clinica). Bull. Soc. Patb. Exot., V, pp. 411-438. 2 plates.
Wi n yon, C. M. (1912). Experiments on the behaviour of Lcishmania and Allied Flagellates
in Bugs and Fleas, with some Remarks on Previous Work. Journ. Lond. Scb. Trap.
Med., II, pp. 13-26.
1
Volume IX
July. 1915
No. 3
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor WARRINGTON YORKE, M.D.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool.
C. Tinting Co., Ltd.
Printers to the University Press of Liverpool
53 Victoria Street
3.49
NOTES ON THE BIONOMICS OF
GLOSSINA PALPALIS IN SIERRA
LEONE, WITH SPECIAL REFERENCE
TO ITS PUPAL HABITATS
[Being the First Report of the Thirty-second. Expedition of the
Liverpool School of Tropical Medicine, 1914-1915]
BY
WARRINGTON YORKE
AND
B. BLACKLOCK
(Received for publication 21 April, 1915)
Plates XXVII-XXXIII and Map
INTRODUCTION
The Thirty-second Expedition of the Liverpool School of
Tropical Medicine was dispatched to Sierra Leone on November
18th, 1914, and returned to England on April 2nd, 1915.
The primary object of this expedition was to find a site suitable
for the permanent research laboratory which the School proposes to
erect in Sierra Leone. Thanks to the kindness of the Administration,
we were enabled to approach the subject under the most favourable
circumstances and to obtain information of the local conditions as
regards population and incidence of disease. After other factors,
such as healthiness, cost of erection and maintenance of the
laboratory, and accessibility from England, had been considered,
we were in a position to make definite recommendations which were
embodied in a report to the Committee of the School.
During our sojourn in the country we made a survey of the chief
parasitic diseases of man and domestic stock, with a view to
ascertaining the lines along which research could with most
advantage be directed in the laboratory which it is hoped to
establish.
The scientific research undertaken by us deals chiefly with the
bionomics of Glossina palpalis. The results of this and other work
accomplished will be published in a series of papers in these Annals.
We desire to express our deep indebtedness to His Excellency
35 °
the Governor, Sir Edward Merewether, to the Principal Medical
Officer, Dr. T. E. Rice, and to the Senior Sanitary Officer,
Dr. R. H. Kennan, for the invariable kindness and courtesy shown
to us during our visit to the Colony, and for the assistance rendered
on every possible occasion.
Some four or five miles from Freetown, at the mouth of the
Sierra Leone River, lies the Cape Lighthouse Peninsula. This
triangular piece of land, of which the long axis is approximately
north and south, is separated from the mainland by Aberdeen Creek.
On the other two sides the Peninsula is bounded by the sea. It is
roughly three miles long, by one and a half miles broad at its base,
with an area of about threequarters of a square mile. It tapers
rapidly and runs as a narrow strip about ioo yards wide for nearly
two miles, until it joins the mainland at its southern extremity. The
Creek is fringed by a dense growth of mangroves, which in certain
places is as much as half a mile broad. On the northern side the
shore is rocky, and on the western side there is a gently sloping
beach of fine white sand (see map).
While oil palms (Eloeis guineensis ) are found all over the
Peninsula, in certain parts they occur exclusively, the appearance of
dense undergrowth being due entirely to the presence of innumerable
young palms (PI. XXVII). In such places which are found especially
in the region adjacent to the mangroves fringing Aberdeen Creek, and
in that portion of the Peninsula on which the lighthouse and isolation
camp are situated, there is deep shade, and little or no long grass.
Between Aberdeen and the isolation camp the growth of palms is
not so dense, and consequently the shade is less; in this locality
• other trees are found, and there is a considerable amount of long
grass. Between the isolation camp and Man of War Bay is a small
area in which oil palms, although present in considerable numbers,
do not predominate to the same extent as elsewhere, many other
trees being present together with a dense undergrowth of shrub.
Besides oil palms, cocoanut palms, mangoes and cotton trees are
represented; there are also a few baobabs, and in the village of
Aberdeen bread-fruit trees, guavas, plantains and golden plum
trees.
Annals Trop. Med. & ParasitolVol. IX
PLATE XXVII
Dense growth of young oil palms.
To face p. 350.
C. Tinlinz < 5 ^ CoLtd., Imp.
Annals Trop. Med. £•? Parasitol., Vol. IX
PLATE XXV111
Hollow baobab constituting a breeding ground of Glossina palpalis.
To face p. 351
C. Tinling & Co., Ltd., l*f-
35 1
The population of the Peninsula is fairly large; in the town of
Aberdeen are some 500 inhabitants, whilst at the Lighthouse and
isolation camp are 50 more, and in addition there are a few natives
living in isolated dwellings. Where the Peninsula joins the main¬
land is the native town of Lumley, with over 400 inhabitants.
Goats, sheep, dogs, domestic fowls and a solitary cow were
found, apparently in good health. Regarding the wild fauna of
the Peninsula nothing very definite is known; monkeys
(Cercopithecus sp.), squirrels, bats, rats and mice are numerous, as
are also many varieties of birds, including bush fowl, guinea-fowl,
ducks and sea birds. A few antelope, bush buck and duiker, were
seen, but they are scarce.
The only fresh water on the Peninsula, apart from the wells
themselves, is the practically insignificant overflow from two or three
springs.
At the beginning of the dry season—December and January—
the tsetse-fly Glossina palpalis was found in fair numbers all over
the Peninsula. A similar prevalence of this fly is recorded by
Dr. W. A. Young, who had spent a month in the locality during the
previous rainy season.
Owing to the circumscribed nature of the Peninsula, its relatively
small size, its proximity to Freetown, and to the fact that cases of
sleeping sickness have been found in the vicinity, the place seemed
well adapted for studying the bionomics of Glossina palpalis. We
were compelled to confine ourselves to one or two of the more
important aspects of the question, as the time at our disposal was
limited.
Although Glossina palpalis is known to be widely distributed
over the West Coast of Africa, and in many places the distribution
of the fly has been carefully mapped out, but little is known of its
pupal habitats in this region. We decided, therefore, to commence
our investigations by a thorough search for pupae. Curiously
enough our earliest effort in this direction was successful, as in the
first place examined no less than eighteen empty pupal cases were
discovered. The site (PI. XXVIII) was a large cavity in the trunk
of a baobab tree. The cavity was conical in shape, having the ground
as its base; the apex of the cone was some 12 to 14 feet above the
ground. The entrance to this chamber was a large triangular
352
opening, about 4 feet wide at the base, which was on a level with
the ground, and about 7 feet high at its apex; it faced south-west.
Owing to the proximity of oil palms and a large mango, no direct
sunlight entered the cavity through this aperture after about
8.30 a.m. One or two small slit-like apertures, about 3 or 4 feet
from the ground, admitted a little light to the chamber from its
northern aspect. The floor of the cavity was, as already mentioned,
flush with the ground, and was composed of a fine dry laterite
gravel covered with dead leaves and debris from the inner surface
of the tree trunk, which was ragged and rugose. The pupal cases
were found after removal of the dead leaves, lying either superficially
or within half an inch of the surface of the ground. Pupal cases
were not found in the numerous crevices of the tree trunk itself, many
of which contained earth probably carried there by ants.
This breeding place was about six yards from the main path
leading to the lighthouse, and about 200 yards from the latter. It
was at least 100 yards from the sea, and not less than a quarter of
a mile from the nearest fresh water, which consisted of a small well.
It is interesting, and probably important, to note that pupal cases
were only found within a foot of the trunk of the tree, and not in
the centre of the floor of the cavity; the explanation of this peculiar
distribution will be discussed later.
Many other places were then searched, but with little success.
These included the intervals between the buttresses at the roots of
cotton trees, dark crevices in rocks, and rock surface densely shaded
by trees, the ground immediately under and overshadowed by fallen
trees and boulders, the cavities at the top of stumps of trees which
had been felled, the sand along the seashore and many other places
of a similar nature. A solitary empty pupal case was discovered on
the sand (PI. XXIX, fig. 1) at the base of a palm tree growing on the
margin of the sandy shore, and another was discovered lying on the
ground in deep shade at the base of a similar tree situated in dense
bush.
We then decided to make a systematic search for pupae on and
around oil palms. With this object we chose at random a young
f palm (PI. XXXI); the tree selected was within 15 yards of the road
and about 200 yards of the lighthouse. It was surrounded and
overshadowed by other oil palms, which in this region formed about
Fig. 2. General view of palm country.
To face p. 352.
C. Tinling <5r* Co., Ltd., Imp
PLATE XXIX
A finals Trop . Med. CS? Parasitol To/. / A'
353
95 per cent, of all the trees present. The trunk of an oil palm
which has its lower petioles still unremoved, as is the case with most
of the palms in the Peninsula, is by no means easy of approach. An
impenetrable barrier is formed by the lower petioles which project
out horizontally in all directions to a distance of 3 feet or more from
the trunk. Frequently the tips of the lowest petioles dip into the
ground, an arrangement which in conjunction with the fact that the
petioles are armed with strong sharp spines effectually prevents any
close examination of the ground in the immediate vicinity of the
trunk of the tree (PI. XXX and XXXI). Having first
searched the ground beyond the range of the petioles and
as far as possible under them without success, the petioles
one by one were cut off at their place of origin from the trunk.
The ground under each was then carefully searched. After
removal of the dead leaves and light debris twenty empty pupal
cases were discovered. The cases were found all round the
trunk, lying, as a rule, not more than 12 inches from it; in every
instance they were on the surface. The ground was dry and sandy,
and covered with a certain amount of fine laterite gravel. The
shade was dense, being derived not only from the petioles of the
palm itself, but also from the close proximity of a large number of
other palms, which almost completely cut off direct sunlight during
the entire day.
The examination of the soil around the trunks of the trees was
greatly facilitated by the use of a spade and newspaper. A
shovelful of superficial soil was placed on the newspaper and spread
out in a thin layer, when the puparia could readily be seen. By this
means the soil at different depths was more accurately and
expeditiously examined. The superficial layer to the depth of
about half an inch was first removed and examined, and subsequently
the deeper layers. It was thus shown that by far the majority
of puparia were either actually on the surface of the ground
or less than half an inch below the surface; the deeper layers
contained very few pupal cases. The following observation
illustrates this point. A palm having been stripped of all its lower
petioles, the ground around it was cleared of dead leaves and twigs.
The soil to an extent of about one yard from the trunk was then
removed in layers and carefully examined for puparia. The
354
following was the result: —The most superficial half-inch contained
fifty-one puparia. The soil between one-half inch and three inches
in depth contained ten, that between three inches and six inches only
two; whilst below this depth no puparia were discovered.
Adopting this procedure, we had not much difficulty in finding
large numbers of puparia of Glossina palpalis in similar sites.
Frequently between ten and twenty pupal cases were taken under a
single palm, and on one or two occasions between twenty and thirty,
whilst in two instances seventy-three and seventy-five puparia
respectively were collected.
As a result of many observations of this kind, we are enabled
to state, as regards this Peninsula, that under any oil palm which has
not had its lower petioles removed, which stands in dense shade and
the ground under which is dry and not too stony, a search for the
puparia of Glossina palpalis would in all probability be successful.
We do not mean to imply by this that, other things being equal,
pupae do not exist where the ground is stony, but simply that they
are more difficult to recognise on such a surface. We put this
postulate to the test by examining a single spadeful of superficial
earth removed from under the petioles close to the trunk of each of
twenty oil palms in a region where practically every tree was a palm.
In two instances only did we fail to find the puparia of Glossina
palpalis by this means.
The angles which the petioles form with the trunk of the palm
contain considerable quantities of earth and debris. These forks
appear to be suitable for the deposition of larvae, and accordingly
a very careful search for puparia was made. Before examining the
contents of the angles, all the pupae lying on the ground to a depth
of three inches were first removed with the aid of a spade, care being
taken to disturb the petioles as little as possible. The ground
around the trunk was then covered with newspapers and the petioles
stripped from the tree, and all the soil and debris in the angles
collected and examined. In this manner several palms were entirely
stripped of petioles, whilst odd petioles were removed from many
other palms (PI. XXXI and XXXII). The result was striking,
a solitary empty pupal case only being discovered in this situation.
At the base of other trees, especially cotton trees and mangoes,
are what appear ideal pupal habitats from the point of view of shade
and soil. The result of searching these places was, however,
Annals Trop. Med. & ParasitolVol. IX
PLATE XXXI
Oil palms ; the lower petioles have been stripped from the tree on the left.
To /ace p. 354 .
C. T in ling Co., Ltd., Imp .
Annals Prop. Med. & Parasitol., Vol. IX
PLATE XXXII
Young oil palm from which all the petioles on the right side had been stripped.
To face p . 354 .
C. Tittling <5r* Co., Ltd., Imp.
PAGE NOT
AVAILABLE
355
decidedly disappointing, only an occasional puparium being
discovered.
As might be expected, most of the puparia taken were empty.
Out of a total of about 450 only twenty were unhatched.* From a
number of these Glossina palpalis emerged after they had been kept
in the laboratory some days.
We have already referred to the fact that the Aberdeen Creek
side of the Peninsula is fringed with extensive mangrove swamp. It
is popularly supposed that there is some connection between the
presence of these swamps and the occurrence of Glossina palpalis.
In fact, the local name for the ‘ fly ’ amongst the natives is ' mangrove
fly.* That portion of the swamp in the vicinity of Aberdeen village
was carefully examined by us firstly for Glossina palpalis , and
secondly for its pupae. It should be mentioned that while the roots
of the mangroves are covered at high tide to a depth varying from
a few inches to three or four feet, at low tide the sea recedes to a
considerable distance beyond their outer fringe, leaving amongst
them small collections of water and tiny streams, so that the
mangroves are seen to be growing in soft sandy mud and pools of
water (PI. XXXIII). An inspection of the mangroves was made on
severed occasions, both at low and high water. The outer edge was
examined on foot at low water, but a boat was necessary at
high tide.
Glossina was found not only in the midst of the mangroves, but
also at their outermost fringe, both at low and high water; this is
of considerable interest, as at certain points where the tsetse was
seen the distance from the dry land was at least half a mile. No
trees were found growing amongst the mangroves below high water,
although just above this point a few mangroves were noticed inter¬
mingled with the ordinary bush growth. The mangroves themselves
afforded fairly good shade, and this, together with the fact that a
considerable number of paths were cut through them to the fishing
grounds beyond, and that the mangroves are inhabited by large
numbers of small birds, mud fish, crabs, shellfish, snails, wading
birds, etc., which may afford a supply of food for the tsetse, is
probably sufficient reason for its presence.
The next question to be considered is, do the mangroves
* The extreme length and breadth of 16 unhatched pupae were measured. The following is
a summary of the results :—Length : max. 6*i mm., min. 5*6 mm; average 5*93 mm. Breadth :
max. 3*6 mm; min. 3*0 mm ; average 3*25 mm.
356
constitute a breeding ground for Glossina palpalis ? Bagshawe
(1911) writes: ‘It is probably lost labour to look in moist soil and
mud for pupae of tsetse flies; they have never been found in any
situation which is not at least moderately dry.* Nevertheless, in
view of the fact that the fly was found amongst the mangroves, we
determined to investigate the point carefully. At low tide, with the
aid of six assistants, all of whom had had considerable experience
in finding pupae, we made an exhaustive examination of the sandy
mud amongst the mangroves and also of the mangroves themselves.
The latter, however, afford very few possible places for the lodgment
of pupae, as the stems are smooth and narrow and do not present
many forks or clefts in which pupae could lodge. Similar examina¬
tions were made at high tide over the same area. No puparia were
found on the mud or floating on the water amongst the mangroves,
nor were any discovered on the stems and branches.
As a result of this work, it appears clear that the tsetse which we
observed in the mangrove swamps were not breeding there, but had
travelled from the land. Support to this conclusion was furnished
by examination of the bush immediately above high-water mark.
Puparia were found there under oil palms in large numbers, and a
few under other trees. Apart from the failure to discover puparia in
the mangrove swamps such places do not appear to be suitable
breeding grounds for tsetse-fly, as pupae which are deposited on
sea-water or on ground which is covered by sea-water for a certain
number of hours each day do not hatch, as is shown by the following
experiment: —
Experiment. —Eighteen unhatched pupae found at the base of oil palms
were placed in glass jars containing a small quantity of sand. Nine of the jars
were half filled with sea-water for a period of three hours daily during a month.
The pupae were observed to float horizontally on the surface of the water, the
tubercles and stigmata being as a rule just below the surface. When the water was
decanted at the end of the three hours, the pupae were left lying on the moist
sand. This experiment was an attempt to imitate the conditions to which pupae
deposited in mangrove swamp would be subjected, except that in the experiment
they were exposed to water once a day only, whereas in nature the tides cover the
mangrove swamps twice daily. At the end of a month the daily addition of water
was discontinued and the pupae were left lying in the sand which soon became
dry. No water was added to the other nine jars, the pupae in which served as
controls. Six of the nine control pupae hatched within the time during which
the experiment was continued, viz. : 35-60 days, depending on the date when the
pupae were discovered. The flies emerged on the 1st, 10th, 20th, 23rd, 27th and
32nd days respectively after the commencement of the experiment. No flies
emerged from the pupae which had been exposed to sea-water. The temperature
was that of the laboratory 8o°-86° F.
357
Carpenter's (1912) work on this subject is of great interest; he
writes ‘According to my experience pupae are only found in the
driest possible situation, though always close to the water’s edge;
so that it would appear that contact with water is inimical to them.
It will be seen, however, that they have a considerable capacity to
resist the effects of submersion; moreover, they do not become mouldy
(so long as they are alive) when kept in an atmosphere with the
maximum degree of humidity. Their powers of resistance to sun
are less in proportion.* He concludes from experiments that should
any breeding ground be flooded at intervals by heavy rains the
conditions would not completely destroy all the pupae. Even four
successive submersions for twenty-four hours destroyed only half the
number. Further experiments showed that it is rather the frequency
of submersion than the total time that is adverse to the pupae. He
states that a certain proportion can survive flotation for eight days,
but none for ten days.
A general survey of the pupal habitats discovered by us in the
Cape Lighthouse Peninsula reveals several important facts.
Firstly, fly and pupae are distributed more or less evenly over
the whole extent of the Peninsula. They are certainly not limited
to the immediate vicinity of water, both being found at least a
quarter of a mile from the seashore. This is the more remarkable
in view of the almost complete absence of fresh water. There is,
so far as we are aware, no record in the literature of palpdlis pupae
being found in any situation except on the water's edge. Bagshawe
(1908), who was the first to discover the pupae of Glosstna palpalis
in nature, writes ‘ One may for the present say that the larvae are
dropped in shade, it may be of shrubs, it may be of bananas, within
forty-five yards of water.' Fraser and Marshall (1909) state that
the distance from high-water mark at which deposits were found was
never more than fifteen yards, the usual distance being five yards;
whilst Carpenter writes * According to my experience pupae are only
found in the driest possible situations though always close to the
water's edge.’ It must be noted, however, that these statements
refer to East Africa. The observers quoted worked on Lake
Victoria Nyanza, where conditions appear to be rather different from
those obtaining in Sierra Leone. Whilst enormous numbers of flies
are to be found in the former place, and they appear to be strictly
358
limited in their distribution to the lake shore and larger rivers, as
regards the Cape Lighthouse Peninsula this is certainly not the case.
Glossina palpalis is not nearly so numerous, nor is it limited to the
immediate vicinity of water.
Secondly, pupae were concentrated on the earth close to the
trunks of trees, and were not found to any extent on the trees
themselves or on the ground between them. By far the majority of
the puparia taken by us were less than one foot from a tree trunk.
The explanation of this appears to be that the tsetse deposits its
larvae in the most secluded and shady spot available. Possibly the
larva is deposited whilst the fly is resting on the trunk or on the
under surface of a lower petiole. Such a hypothesis would account
for the peculiar distribution of the puparia in the hollow baobab
referred to previously. Tsetses resting on the inner surface of the
hollow trunk would drop their larvae on the ground close to the tree
trunk, thus explaining the peripheral distribution of the puparia on
the floor of the chamber. Similarly in the case of the oil palm, flies
resting on the under surface of the lower petioles or on the tree trunk
would also drop their larvae on the ground near the trunk.
The facts discovered by Moiser (1912) are of interest in this
connection. He put men in trees at a height varying from 10 to
25 feet for an hour; not one of them reported having seen a fly,
though several ( Glossina tachinoides') were seen on the ground during
the period. In order to determine where flies usually rest, he put
up a large mosquito net (8 x 8 x 8 ft.) in the bush, thus enclosing
a portion of the natural haunt of the fly. Eleven Glossina
tachinoides were then liberated within the net. A number of them
quickly disappeared from view, and after a few minutes* search he
discovered them resting in an inverted position on the under side of
small branches and twigs close to the ground. Branches between
6 inches and 2 feet from the ground were those most frequently
occupied by the tsetses, and in order to observe them closely Moiser
had to lie on the ground. Subsequently flies were found at rest on
the under surface of small branches outside the net.
Thirdly, the ground at the base of oil palms appears to be a most
suitable breeding place for Glossina palpalis. In this region
puparia of Glossina palpalis can be found under almost any oil palm
which has not had its lower petioles removed, which stands in dense
359
shade, and the ground under which is dry and not too stony.
Moreover, Glossina palpalis is able to breed in a locality where oil
palms of various sizes and ages 'afford the sole shelter. The lower
surface of a petiole is broad and smooth and well protected from
the sun, and tsetse have been observed in this situation. A fly in
this position is hidden from view except to anything actually on the
ground, and is also to an extent protected from molestation by the
sharp strong spines with which the petioles are armed. In so far
as the Cape Lighthouse Peninsula is concerned, the situations in
which pupae are deposited are hidden from view, probably as a
protective measure for the fly undergoing parturition and for the
newly deposited larva against birds, a view which would explain
the absence of puparia in many spots, such as the spaces between
the buttresses of the trunks of cotton trees, otherwise apparently
well adapted as pupal habitats.
Zupitza (1908), working at Duala in the Cameroons, recorded
that he had found the puparia of Glossina palpalis in the humus and
moss in the forks of branches and in the cracks in the bark of all
trees, especially in the angles of leaf sheaths of palms at a height of
a few centimetres to three and a half metres above the ground. He
did not look for them at a higher level. They were never found in
the dry mould of hollow trees, nor in or on the ground. The
observations of Zupitza are in curious contradistinction to those
made by us. The first pupal habitat discovered by us was the
ground forming the floor of the chamber in the baobab trunk.
Moreover, whilst we constantly found puparia on the ground close
to the trunk of oil palms, only a single empty case was discovered
in the angles between the petioles and the tree trunk. As already
mentioned, we most carefully investigated this place as a possible
pupal habitat, entirely stripping the petioles from three or four
palms on the ground under which large numbers of pupae had
been found, and examining the debris between them and the tree
trunks without finding any pupae. In addition, odd petioles were
stripped from many other palms with equally negative results.
The explanation of this apparent discrepancy is cryptic. We are
not able to suggest any reason why Glossina palpalis should in one
locality deposit its pupae on the ground and in another on the tree
itself, unless in the latter case the ground happened, through
3 < 5 o
moisture or some other unknown condition, to be an unsuitable pupal
habitat. Zupitza states that in the angles between the petioles and
the trunks of the palms the pupae would be kept moist, but that they
would be in no danger of being drowned or washed away by rain¬
water running down the trunk. He is evidently under the
impression that some amount of moisture is necessary, as he never
found them in the dry moss in the holes of trees, nor in or on the
ground. This is not our experience, as they were almost invariably
found lying on or just under the surface of perfectly dry ground.
Before leaving this subject, it is of interest to consider what steps
it would be necessary to take in order to clear the Peninsula of
tsetse. In view of the fact that the fly is ubiquitous in the area under
consideration and that its pupae have a like distribution, it is quite
obvious that no localised or partial clearing could be recommended
as likely to eradicate the fly. Moreover, in districts where practi¬
cally all the vegetation consists of oil palms, wholesale removal of
these cannot, for economic reasons, be contemplated. Yet we are
faced with the fact that it is under the young palms that the tsetse-
fly find their best breeding place; any method to be practicable
must not interfere with the economic value of the trees. The point
that has to be considered, therefore, is whether any measures
short of actual destruction of the trees would suffice to
get rid of the fly. In our experience the ground at the
base of an oil palm which has had its trunk cleared of petioles
does not constitute a breeding place of palpalis. The soil
at the base of many such trees was searched without success.
We are convinced that it is the presence of the lower petioles that
renders young oil palms such excellent breeding places. Removal
of these would, in our opinion, destroy the breeding grounds of
palpalis , and by this means such localities could be freed of fly
without loss by damage to the trees. Such a procedure would be
laborious, but it must be remembered that the petioles when once
removed do not grow again. In old palms the lower petioles are
absent; as the tree grows the lower, that is the older, petioles become
dry and rotten and are easily stripped off, but in very young
palms they are much tougher, and hence more difficult to remove.
Naturally, in those places where shrub growth, other than young oil
palms, occurred, it would have to be removed.
361
It may not be considered advisable on economic or epidemio¬
logical grounds to attempt to rid the Peninsula of tsetse at the
present time. If, however, this can be undertaken as an experiment,
results of the utmost value would be obtained, and the information
gained could be applied in districts where for economic or
epidemiological reasons it might be of vital importance to exterminate
Glossina palpalis. Owing to its proximity to Freetown, its small
size and its well circumscribed character, the Cape Lighthouse
Peninsula is eminently suitable for an experiment of this nature.
CONCLUSIONS
1. The breeding grounds of Glossina palpalis are not so
strictly limited to the immediate vicinity of water as has hitherto
been thought; they may occur quite independently of fresh water
and at least a quarter of a mile from sea water.
2. Although Glossina palpalis is to be found in considerable
numbers in mangrove swamps and may travel in these to a distance
of at least half a mile from dry land, the swamps do not constitute
a breeding ground of the fly.
3. The pupae of Glossina palpalis do not hatch when subjected
to daily flotation on sea water.
4. The ground around the trunk of oil palms ( Eloeis guineensis)
which have not been stripped of their lower petioles constitutes an
excellent breeding place for Glossina palpalis .
5. Glossina palpalis can breed in localities in which practically
the only tree is the oil palm.
6. Stripping the oil palm of the lower petioles would suffice to
destroy the breeding ground in such localities.
REFERENCES
Bagshawe, A. G. ( i 911). Sleeping Sickness Bulletin , Vol. Ill, p. 362.
- (1908). Reports of the Sleeping Sickness Commission of the Royal Society , No. IV, p. 48.
Carpenter, G. D. H. (1912). Progress Report of Investigations into the Bionomics of Glossina
palpalis , July 27, 1910, to August 5, 1911. Report of the Sleeping Sickness Commission of
the Royal Society } No. XII, pp. 79-1 n.
Frazer and Marshall (1909). Progress Reports on Uganda Sleeping Sickness Camps. London
Sleeping Sickness Bureau , pp. 40-44.
Moiser, B. (1912). Notes on the haunts and habits of Glossina palpalis. Bull. Ent. Res Vol.
Ill, pp. 195-202.
Zupitza, — (1908). Ueber die Schlafkrankheitsfliege bei Duala. Beibefte zum Arcbiv. f. Scbiff.
und Tropen. Hyg. y B. 12, pp. 1-27.
J63
FOOD OF GLOSSINA PALPALIS IN
THE CAPE LIGHTHOUSE PENINSULA,
>> SIERRA LEONE
[Being the Second Report of the Thirty-second Expedition of the
Liverpool School of Tropical Medicine, 1914-1915.]
BY
WARRINGTON YORKE
AND
B. BLACKLOCK
(Received for publication 28 April , 1915)
Although there is a certain amount of evidence to show that
Glossina palpalis can take up water and vegetable juices, practically
all observations indicate that for its continued existence vertebrate
blood is necessary. In view of the small amount of time at our
disposal, and of the comparative scarcity of Glossina palpalis , we
were able to examine only 200 flies in respect of the nature of the
blood found in their intestinal tract. The tsetse were dissected
immediately they were brought into the laboratory. As at the same
time the flies were examined for the presence of trypanosomes, they
were dissected in the manner described by Lloyd (1912). Briefly
it consists in splitting the dorsum of the thorax longitudinally with
a cataract knife, and then drawing out the salivary glands by gentle
traction on the head. Usually the glands are removed intact
attached to the head, the oesophagus breaking off at the point where
it enters the pharynx; occasionally they break across when only
partially withdrawn, but in this case are easily caught up with a
fine pair of forceps. The proventriculus is then sought at the ventral
surface of the thorax, and is drawn out with the sucking stomach
and most of the intestine. The posterior portion of the hind-gut
and rectum are removed by cutting off the last segment of the
abdomen. The gut contents and those of the salivary glands and
of the proboscis were examined between a slide and coverslip in a
fresh unstained condition.
364
Recognisable red blood cells were seen in 16 of the 200 flies thus
examined. It is necessary to point out here that only those flies in
which definite red blood corpuscles were seen are recorded as
containing blood. Practically every fly had in its mid-gut a certain
quantity of pigmented material, the colour of which varied from
bright red to brown or black. Such pigmented material does not,
however, necessarily imply the presence of red blood corpuscles; on
microscopical examination it was usually found to consist of granular
debris and globules appearing red or brown in colour, and
crystals which in many cases were probably haemin or haemoglobin
crystals. Although it is highly probable that this pigmented
material was derived from blood, still one could not be certain, and,
therefore, in the following table only those flies in which definite
red blood corpuscles were encountered are considered as containing
blood.
Tablx I.—Result of examination of freshly-caught wild Glonima fslptlts for recognisable blood
corpuscles
Number
examined
Number in which
recognisable
mammalian red
corpuscles were seen
Number in which
recognisable
nucleated red
corpuscles were seen
Males .
113
8
0
Females .
!
87
6
2
I
Total .
200
14
2
From the limited number of observations made, it appears that
eight per cent, of freshly caught Glossina palpalis in this Peninsula
contain recognisable red blood corpuscles—mammalian blood in
seven per cent., and nucleated red cells of an undetermined nature
in one per cent. The chief source of mammalian blood on the
Peninsula is probably man and his domestic stock—goats, sheep,
and dogs. In addition there are a few antelope—bushbuck and
duiker were seen—and a considerable number of monkeys
(<Cercopithecus sp.) squirrels and bats, rats and mice. It is
interesting to note that the mammalian red cells seen in the fourteen
flies observed to contain these, were of the large type and readily
distinguishable from the small variety found in sheep and goats.
36s
The source of nucleated blood corpuscles is apparently much
larger. Birds and lizards are numerous, and in the mangrove
swamps there are vast numbers of mud-fish, crabs, shell-fish, snails,
wading birds and sea birds. No crocodiles occur. In view of the
fact that the reservoir of nucleated blood corpuscles is probably much
greater than that of non-nucleated red cells, it is interesting to find
mammalian blood in fourteen Glossina palpalis and nucleated red
cells in only two, an observation which appears to indicate that
Glossina palpalis either prefers mammalian blood or finds it more
easy to obtain. The flies were captured at various parts of the
Peninsula, but owing to its small size none of these localities was
more than a quarter of a mile from human habitation—either the
village of Aberdeen or the settlements at the isolation camp and the
Cape Lighthouse.
In order to appreciate the real meaning of the results obtained
by such an examination of the gut of freshly caught tsetse in respect
of blood corpuscles, it is of importance to have at our disposal some
data regarding the length of time red blood cells can be recognised
in the intestine of a fly after feeding. To obtain this information
freshly caught tsetse were fed on a rat or fowl and dissected after
various intervals, and the gut contents examined for the presence of
red blood cells. The results are given in the Tables II and III.
In the case of mammalian blood it is seen that in over 90 per cent,
of the flies red cells can be recognised 24 hours after a feed, but in
only 40 per cent, of those examined after 48 hours, whilst in no
instance were definite red cells noted after 72 hours. The fowl red
blood cells were found to be recognisable for longer periods; after
24 hours 100 per cent, of the flies showed nucleated red blood cells,
whilst after 48 hours red blood cells were seen in 60 per cent., and
after 72 hours in 40 per cent. The temperature was that of the
laboratory, 8o°-86° F. Possibly to a certain extent this difference
is to be explained by the fact that the nucleated red blood
cells of the fowl are more characteristic, and hence more easily
recognised than are the non-nucleated red cells of mammals. Of
course, these figures are approximate, as only 64 tsetse were
employed in the experiment: they are, however, sufficiently accurate
for practical purposes, although some observers have recorded, in the
case of isolated flies, that blood could be found after much longer
3 66
intervals. Thus in the case of Glossina morsitans , Lloyd (1913)
observed fowl red cells in a clot in the sucking stomach several
weeks after the fly had last fed on a fowl, monkeys having been used
as blood donors in the interval. Nevertheless, these figures show
Table II.—Experiment to ascertain the length of time rat red blood corpuscles can be recognised in
G. palpalis
No.
Date fed
Date
examined
No. of
hours
after
feed
Sex
Result of
examination.
Recognisable
blood cells
present or absent
Percentage of flies
in which
blood was found
1
18.12.14
24
<5
+ + +
2
17.12.14
18.12.14
2 4
(J
+ + +
3
17.12.14
18.12.14
24
$
+ + +
4
17.12.14
18.12.14
24
9
+ + +
5
17.12.14
18.12.14
24
9
+ + +
6
17.12.14
18.12.14
24
3
+ + 4 -
9 2
7
17.12.14
18.12.14
24
9
4 - + +
8
18.12.14
24
<?
4 - 4 -
9
2 9- I * , 5
24
<?
—
10
2 9* I * I 5
24
<?
4 - 4 - 4 -
11
9
3* 2 * I 5
24
<?
4 - 4 - 4 -
12
2.2.15
3- 2 * , 5
24
<?
4 - 4 - 4 -
*3
28.1.15
48
_
28.1.15
48
9
—
*5
28.1.15
48
<?
4 - 4 - 4 -
16
28.1.15
1
48
9
—
l 7
28.1.15
48
9
+ 4 - 4 -
40
18
2.2.15
48
9
—
*9
2.2.15
48
<j
—
20
2.2.15
48
9
+
21
2.2.15
. 9
48
—
22
2.2.15
4* 2 * , 5
48
<j
+ + +
2 3
28.1.15
3 »»>S
7 2
—
2 4
28.1.15
3 I-I-I 5
7 2
cf
—
2 5
28.1.15
31.1.15
7 2
9
—
26
2.2.15
5 * 2 * 5
7 2
9
—
2 7
2.2.15
5- 2 *5
7 2
-
28
2.2.15
5- 2I 5
7 2
<?
—
0
2 9
2.2.15
5 * 2 - 5
7 2
9
—
30
2.2.15
5- 2I 5
7 2
9
—
3 *
2.2.15
5- 2, 5
7 2
<?
—
3 2
2.2.15
5* 2<I 5
7 2
9
—
33
2.2.15
5- 2I 5
7 2
—
34
21.1.15
2 5 .i.i 5
96
<?
that as a general rule mammalian (rat) red blood cells are no longer
recognisable after a period of 72 hours, whilst fowl red cells are
observed in only 40 per cent, of cases after a similar period. This
information is of importance in connection with that obtained by
3 67
examining the blood found in freshly caught tsetse-fly. When the
statement is made that only seven per cent, of such flies contain
mammalian blood, it must be borne in mind that if the flies had all
fed on a mammal 48 hours previously, we would expect to find
red cells in only 40 per cent., whilst if they had fed 72 hours
previously very few would contain recognisable red cells.
Tablk III.—Experiment to ascertain the length of time fowl red blood corpuscles can be recognised in
G. palpalis
No.
Date fed
Date
examined
No. of
hours
after
feed
Sex
Result of
examination.
Recognisable
blood cells
present or absent
Percentage of flies
in which
blood was found
1
28.1.15
29.1.15
24
3
+ + +
'I
2
3 II - , 5
24
3
+ + +
3
30.1.15
3 ,II 5
24
6
+ + +
4
30.1.15
3 III 5
24
3
+ + +
5
I.2.I5
2.2.15
24
9
+
100
6
I.2.I5
2.2.15
24
3
+ + +
7
I.2.I5
2.2.15
24
?
+ + +
8
I.2.15
2.2.15
24
3
+ + +
9
1.2.1 5
2.2.15
24
3
+
10
I.2.I5
2.2.15
24
9
+ +
11
28.I.I5
30.1.15
48
3
_
12
28.1.15
30.115
48
?
—
! 3
28.1.15
30.1.15
48
3
—
H
28.I.I5
3°* I * I 5
48
3
+ + +
i 5
28.I.I5
30.1.15
48
$
+ + +
60
16
1-2.15
3.2. 1 5
48
3
+
l 7
I.2.I5
3.2.15
48
3
+ + +
18
I.2.I5
3 - 2-15
48
3
—
>9
I.2.I5
3 - 2 -i 5
48
3
+ + +
20
I.2.I5
3.2.15
48
3
+ + +
21
28.I.I5
3 i-i-i 5
72
3
—
22
28.1.15
3 i-i-i 5
72
3
+
23
28.1.15
3 i-i-i 5
72
?
+
28.1.15
3 i-i-i 5
72
3
-
28.1.15
3 i-i-i 5
72
9
—
40
26
30.1.15
2.2.15
72
3
—
27
30.1.15
2.2.15
72
3
+ + +
28
30.1.15
2.2.15
72
9
—
29
30.I.I5
2.2.15
72
3
+ +
30
30.1.15
2.2.15
72
3
The question whether tsetse-fly take up other food than blood
is one which is difficult to decide. Stuhlmann (1907) and Degen
(1909) came to the conclusion that they did not, but Maugham (1911)
states that he has seen tsetse-flies sucking vegetable juices on two
occasions. In 1905 he observed a Glossina morsitans alight on a
368
stem of young marsh grass ( Phragmites communis) and deliberately
insert its proboscis and unmistakably suck for a period of about
three and a half minutes. At this stage Maugham caught the fly,
and found on examination that it was partly full of the moisture
from the plant. Again in 1908, in an absolutely gameless and
practically waterless country, he observed Glossina morsitans feeding
on a piece of sugar-cane at a point where the pith was exposed. He
attempted to catch this fly, but unfortunately failed to do so.
Taute (1912) investigated the point experimentally in the following
manner. Two hundred tsetse-flies which had been fed daily on
blood for a period of two months were starved for five days, and
subsequently were given an opportunity of feeding on small pieces of
mango fruit. Taute records that in three cases Glossina morsitans
buried its proboscis completely in the fruit and remained in this
position for several minutes, but definite sucking did not take place.
One of the flies was killed and dissected immediately after this
operation. Neither in the lumen of the proboscis nor in the
oesophagus or remainder of the alimentary canal could the smallest
trace of mango juice be recognised.
The work of Carpenter (1912-13), however, affords strong support
to the view that Glossina palpalis does take up other food than
blood. Carpenter examined the intestinal contents of a large number
of freshly caught G . palpalis , and found in a proportion of
them small fragments of tissue of an obviously vegetable origin,
e.g., pieces of vegetable parenchyma, starch grains, pieces of alga,
and a minute fungus.
Apart from the solitary instance recorded by Maugham, there
appears to be no direct evidence that tsetse-fly will imbibe anything
other than vertebrate blood, and, moreover, the evidence offered by
him is by no means convincing, as he omits to state the manner in
which he examined the fly or how he detected that it was partly
filled with moisture from the plant. Stuhlmann affirms that Glossina
lives exclusively on living blood, and that they refuse to take up
shed blood, water or syrups; and Degen attempted to feed them on
fruit, saccharine fluids, meat, etc., without success. Rodhain,
Pons, Vandenbranden and Bequaert (1912) observe that it is
generally admitted that tsetses cannot engorge themselves with
blood unless they obtain it directly from the capillaries in which the
369
fluid is maintained under a definite pressure. Leaving pressure
out of the question for the moment—Rodhain and his collaborators
have since shown that this is not an essential factor—all experimental
evidence shows that Glossina are unable to imbibe shed blood. In
the experiments described below we have demonstrated that they will
not absorb certain other fluids offered to them in open vessels.
Fourteen flies which had been starved for 48 hours were placed in
each of two cages. In one of these was a petri dish filled with the
following solution:—
Sodium chloride. 0*9 g.
Sugar . 2 - og.
Neutral red . o ig.
Water . 100 c.c
and in the other a petri dish containing the following: —
Methylene blue. 0*05 g.
Sodium carbonate . 0*025 S-
Water . 100 c.c.
Small twigs and leaves were floated on the surface of the solutions.
The flies were dissected as they died; all those that were still alive
at the end of 48 hours were killed and examined. There was no
indication that any of the flies had imbibed fluid. These results
are conclusive because, as will be seen later, solutions of neutral red
and of methylene blue in the concentrations used, if taken up by
Glossina, stain the tissues densely. Such work as we have done,
therefore, confirms the generally accepted view that Glossina is
unable to imbibe exposed fluids directly.
The work of Rodhain and his collaborators showed, however,
that tsetses can suck up citrated blood through a membrane consisting
of the freshly removed skin of a mouse. We made use of this
observation to prove that not only will tsetses take up citrated blood
through such a membrane, but that they will also imbibe various
other fluids. The apparatus used by us was a slight modification of
that figured and described by Rodhain. It consisted of a short glass
cylinder about three-quarters of an inch in diameter, the lower end
of which was closed by a cork through which the shorter limb of an
b-shaped piece of glass tubing passed; the upper end of the
cylinder was covered by the membrane. The pressure of the fluid
in the cylinder was indicated by its level in the longer limb of the
glass tube.
37 °
In our first series o£ experiments flies were fed on defibrinated
goat-blood through a membrane of fresh rat’s skin. It was observed
that (within the limits of the experiment) the pressure made no
difference to the manner in which the flies engorged themselves.
They distended themselves without the least difficulty at pressures
ranging between + or — 60 mm. of blood. It was further noted
that full distension of the flies occurred at least as rapidly and
regularly as when the flies were allowed to feed on live rats. On the
Table IV.—Giving the results of feeding G. palp alts on defibrinated blood and various dilutions of this with
normal salt solution, through a* membrane consisting of rat skin
No.
Sex
Membrane
Nature of fluid
Result
Remarks
,
<?
Rat skin .
Undiluted goat blood
Complete distension
Immediately
2
n
it
11
3
9
a
a
a
4
9
11
a
it
11
5
3
it
a
ii
a
6
9
M
a
ii
a
7
<?
II
50% goat blood
it
a
8
3
it
n
it
a
9
9
it
n
a
a
IO
3
»
25% goat blood
a
a
ii
9
n
i)
it
a
12
3
it ••• •••
a
a
a
*3
3
11
10 % £ oat blood
a
a
*4
9
a
n
Partial distension ...
After 15 minutes
>5
9
tt
a
11
i»
16
3
a
10% rat blood
Complete distension
Immediately
*7
3
a
a
a
11
18
3
a
a
a
11
19
3
a
5% goat blood
No visible distension
After 15 minutes goat*'
cells seen in anterior gut
20
9
a •••
11
1*
After 1 5 minutes no goats’
cells seen in anterior gut
21
9
a
5 % rat blood.
11
After 15 minutes rats’ cell*
seen in anterior gut
22
9
jj ••• •••
n
Partial distension ...
After 15 minutes
^3
3
ii
No visible distension
After 15 minutes red cells
seen in anterior gut
24
3
ii
a
Marked distension ...
After 15 minutes
other hand the flies attacked the membrane with less alacrity than
they do the skin of the living animal. Nevertheless, many flies
settled on and pierced the membrane at once, and, as a rule, a little
patience was sufficient to induce a large proportion of the flies to
feed. An experiment was conducted to determine the effect of
diluting the defibrinated blood with salt solution in various degrees;
the results are given in Table IV. It was found that Glossina palpalis
37i
engorged itself with the following dilutions, viz., 50 per cent, blood,
25 per cent, blood, and 10 per cent, blood, but the dilution
consisting of 5 per cent., blood and 95 per cent, salt solution was
not taken up by the tsetses with the same rapidity and regularity
as the other solutions. Of the six flies which were offered the last
dilution of blood, only two partially distended themselves; in the
other four no distension was observed, but in three of these red ™»11c
were seen on dissection.
Physiological salt solution alone was then tried, but although the
flies attacked the membrane with the same eagerness as before, no
visible distension was noted, except in one instance where the fly
appeared to become partially engorged. Tsetses frequently inserted
their proboscis and appeared to endeavour to feed; in some
instances the membrane was pierced probably at least 100 times with
apparently no result. We shall return later to the question whether
these flies had actually taken up any salt solution, and simply
note here that no distension comparable to that observed when
deflbrinated blood is offered took place.
We next turned our attention to the question of what element of
the blood proves so attractive to the fly that it engorges itself with
this fluid to such a marked degree. Fresh deflbrinated goat-blood
was centrifugalised, and the red cells separated from the plasma;
the red cells were then washed free from plasma with normal sodium
chloride solution and a 50 per cent, suspension of them made in
salt solution—a concentration which corresponds approximately to
that of red cells in normal goat-blood. It was found that Glossina
palpalis engorged itself readily with the red cell suspension. By the
addition of normal salt solution, suspensions containing 25 per cent.,
10 per cent, and 5 per cent, of red blood cells were made. The
results of offering these to the flies are given in Table V. The tsetse
engorged themselves completely with all the higher concentrations,
but in the case of the 5 per cent, suspension of red blood cells only
one of three flies became completely distended, whilst in the other
two no distension was observed, although red cells were found in the
gut on dissection.
Attempts were made to feed a number of Glossina palpalis on
fresh deflbrinated plasma. The plasma was obtained by shaking
up goat’s blood in a bottle with beads. After separation of the fibrin
372
by straining through gauze, the red cells were thrown down by
centrifugalisation and the plasma siphoned off. Plasma thus
prepared is always of a slightly reddish tint, owing to a certain
amount of damage to red cells during the process of defibrination,
but the amount of haemoglobin dissolved is so small as to be
Tabu V.—Giving the results of feeding G. palpalis on suspensions of red blood cells of various concentration
through a membrane of rat skin
No.
Sex
Membrane
Nature of suspension
Result
Remarb
i
(?
Rat sldn .
5° % goat erythrocytes
Complete distension
Immediately
2
?
»
ii
11
i>
3
<j
» •••
ii
11
11
4
<?
n
ii
n
1?
5
<j
ii ... ...
ii
11
11
6
$
n
ii
n
11
7
,, .
25 % goat erythrocytes
11
11
8
<?
11
1*
ii
11
9
<?
11
i° % goat erythrocytes
Partial distension ...
After 15 minutes
IO
$
11 •••
>»
>»
s ii
ii
c 1
„ .
u
Complete distension
Immediately
12
<?
„ .
5 % goat erythrocytes
i>
After 15 minutes
>3
9
11 ... ...
11
No visible distension
After 15 minutes goats'
erythrocytes seen in gut
14
<J
11
ii
11
n
Table VI.—Giving the results of feeding G. palpalis on defibrinated plasma through a membrane of rat skin
No.
Sex
1
Membrane
Nature of fluid
Result
Remarb
1
<?
Rat skin .
70 % plasma and 30 %
normal saline
No visible distension
After 15 minutes
2
9 !
>1
11
Slight distension
11
3
<?
n
ii
No visible distension
11
4
<?
ii
11
n
11
5 !
s
n .
11
ii
11
6 i
<?
11
11
Almost complete
distension
11
7
11
11
No visible distension
11
8
9
” .
n
Complete distension
11
practically negligible. Two of eight flies, all of which had made
repeated efforts to feed, succeeded in completely engorging
themselves. In only one of the remaining six was slight distension
noticeable. The results are given in Table VI.
!
l
373
From these experiments it is apparent that the most attractive
element in the blood is the red corpuscle. In order to carry the
matter further, the washed red cells of the goat were laked by the
addition of three parts of distilled water to two parts of red cells.
This solution was offered to three Glossina palpates, all of which
quickly and completely engorged themselves, thus proving that the
integrity of the red corpuscles is not an essential factor. An attempt
was then made to separate the red cell stromata from those
constituents of the corpuscle which are soluble in water. For this
purpose sufficient sodium chloride was added to the laked red cell
solution to render it isotonic, and the stromata were subsequently
precipitated by prolonged centrifugalisation. Unfortunately, we
did not succeed in obtaining complete separation of the stromata, as
we had no high power centrifuge at our disposal, and owing to the
minute size of the goat’s erythrocytes precipitation of the stromata
is a matter of no small difficulty. Nevertheless, although complete
separation was not obtained, a considerable fraction of the total
amount of stromata was precipitated and a solution of goafs
haemoglobin, fairly free from solid matter, was obtained. The
resulting solution contained approximately as much haemoglobin as
that found in a 40 per cent, suspension of goafs red cells.
Four flies to which this solution was offered rapidly and
completely distended themselves. The solution was then diluted
with an equal volume of physiological saline. It was found that the
six flies to which this solution was offered did not feed with the same
avidity as did those which were offered the more concentrated
solution; one became completely distended, three others partially,
whilst in two no distension occurred.
Having thus determined that Glossina palpates feeds readily on
fresh haemoglobin solution, further experiments were performed with
solutions made from the crystallised haemoglobin of commerce
(Griiblefs dried haemoglobin). Solutions made from this prepara¬
tion are of a brownish-red colour, and give rather indistinct
absorption bands of oxyhaemoglobin. The strength of the solution
used was between 1 and 2 per cent. Four of the five flies to
which this solution was offered became partly distended, whilst in
the case of the other, although no distension was visible, the solution
was seen in the gut on dissection. None were found to engorge
374
themselves completely in a manner comparable to that seen with
haemoglobin solution made from fresh blood. Details are given in
Table VII.
Owing to the lack of suitable apparatus, we were unable to
determine the result of offering to Glossina palpalis the washed
stromata derived from red blood cells.
Table VII.—Giving the results of feeding G. palpalis on haemoglobin solutions of various concentration
through a rat skin membrane
No.
Sex
Membrane
Nature of fluid
Result
Remarks
1
?
Rat skin .
40 % # solution of goat
haemoglobin in 0*9 %
NaCl
Complete distension
Immediately
2
a
n
tt
After 5 minutes
3
a
n ••• •••
tt
11
Immediately
4
$
,,
it
tt
it
5
?
tt
20 %• solution of goat
haemoglobin in 0*9 %
NaCl
Partial distension ...
After 15 minutes
6
$
n
tt
Slight distension
it
7
$
tt
n
Almost complete
distension
tt
8
a
11
tt
No visible distension
tt
9
$
» •••
tt
Slight distension
tt
10
a
11
it
No visible distension
tt
11
$
it
1 — 2% solution of dry
crystalline haemoglobin
Partial distension ...
tt
12
a
it
n
it
tt
13
a
1
M
tt
tt
tt
After 15 minutes Hb.
solution seen in anterior
gut
*4
a
It
11
No visible distension
<5
a
n
tt
Slight distension
After 15 minutes
* By this is meant that the solutions contained respectively as much haemoglobin as do 40 and 20 P«
cent, suspensions of goat red blood cells.
Summarising the results of these experiments, we find that
Glossina palpalis feeds with avidity, through rat’s skin, on fresh
defibrinated blood and also on suspensions (50 to 5 per cent.) of
washed red blood cells in normal saline, and on solutions containing
as much dissolved haemoglobin as is present in 40 and 20 per cent,
suspensions of red blood cells. Defibrinated plasma does not appear
to have the same attraction for them, nor does sodium chloride
solution alone, but the latter, containing a small proportion of the
dried haemoglobin of commerce in solution is to a certain extent
375
taken up. Although these experiments are not quite so conclusive—
especially as regards separation of the constituents of the red blood
cells which are soluble and insoluble in water—as could be desired,
nevertheless, we consider they suggest strongly that the element in
the blood which is attractive to Glossina palpalis is that fraction of
the erythrocyte which is soluble in water, most probably haemoglobin.
Having completed these observations on blood, we decided to
make use of the same technique with a view to determining whether
Glossina palpalis will take up other solutions through fresh rat-skin.
Reference has already been made to the fact that when sodium
chloride solution was offered, although the flies repeatedly pierced
the membrane, they did not appear to take up any of the fluid. At
all events, no distension was noticed as a rule, although in one
instance partial engorgement did apparently occur; it was by no
means easy to state definitely whether or not any of the solution had
been imbibed, as small quantities of a clear fluid cannot be
recognised in the gut of the fly. In order to determine the point,
the solution must be coloured with some dye that can be readily
recognised. For this purpose methylene blue, neutral red and
fuchsin were used. It was found that when physiological saline,
containing these dyes in solution, was offered to Glossina palpalis
small quantities were taken up by the flies in a proportion of the
instances. Dissection showed that the intestinal tract was deeply
stained, red or blue according to the dye used. In some cases the
stain had spread to the salivary glands and to the fat bodies. In
fact, so deeply were certain flies coloured that even before dissection
the dye could be recognised through the integument of the abdomen.
After obtaining this information various other solutions were offered
to Glossina palpalis , details of which are given in Table VIII. It is
interesting to note that three flies became completely engorged with
0 9 per cent, sodium chloride solution to which had been added
about 5 per cent, of cane-sugar and a little neutral red. Another fly
became partially distended on a 25 per cent, solution of glycerine
in water coloured with methylene blue.
These observations demonstrate that under certain conditions
Glossina palpalis will imbibe and even completely distend itself
with fluids other than blood. Although in all the experiments just
described the membrane used was the freshly removed skin of a rat
376
or rabbit, nevertheless we found that this tsetse will feed through
other membranes, e.g., sheep’s bladder, peritoneal tissue and
membrane composed of a thin sheet of rubber. As regards the last,
one fly out of five completely distended itself with defibrinated blood
through this membrane. Although the flies had not the slightest
Table VIII.—Giving the result of feeding G. palpalii on various fluids through a membrane of skin
No.
Sex
Membrane
Nature of fluid
Result
Remarks
1
$
Fresh rabbit skin ...
Methylene blue
No visible distension,
Anterior and mid-gut blue.
solution*
but abdomen of
bluish tinge
Posterior gut green.
Salivary glands, mak
pighian tubules and fat
bodies blue
2
<?
99
it
No visible distension
99
3
Same rabbit skin 24
>1
99
No staining seen on di»*
hours old
section
4
c?
99
>>
99
99
5
?
t>
99
Proventriculus and anterior
gut blue
6
cJ
No visible distension,
Whole intestine (except
rectum), salivary glands,
but abdomen of
bluish tinge
malpighian tubules and
fat bodies deeply stained
7
?
99
Neutral red -i% solu-
No visible distension
Anterior gut red
tion in water
8
cf
99
99
99
9
9
Fresh rat skin
Fuchsin *1 % in *9%
Slight distension
Proventriculus and an tenor
NaCl solution
gut pink
10
?
99
t>
99
99
11
£
»
Neutral red • 1 % solu¬
Considerable
Whole gut (except rec¬
tion in water + sugar
distension
tum), salivary glands,
malpighian tubules and
Complete distension
fat bodies deeply stained
12
9
99
>»
! 99
>3
9
»»
99
99 t
i . 99
14
9
99
Methylene blue
No distension
Anterior and mid-gut deep
solution + sugar
blue
15
£
99
99
99
16
£
99
99
99
Slight staining of anterior
gut
*7
<3
,,
75% methylene blue
99
No staining
solution + 25%
glycerine
18
£
99
99
Partial distension ...
Anterior and mid-gut and
also salivary glands and
fat bodies stained
*9
£
99
t>
99
Anterior gut blue
• The methylene blue solution used was the weak solution employed in Romanowsky's stain; its formula
is given on page 369.
difficulty in piercing the rubber, they did not seem able in the
majority of cases to imbibe blood through it; possibly owing to the
extremely elastic nature of the rubber the lumen of the proboscis was
occluded, thus preventing the passage of fluid through it.
377
The practical point that we have to decide is, does Glossina in
nature obtain food other than blood ? Although this question is not
answered by the experiments described above, yet the results
obtained are very suggestive. They show that under favourable
conditions Glossina will take up solutions of vegetable origin, such
as sugar and water. Apparently one essential is that the fluid
should be presented enclosed by a membrane. The reason for this
Tabls IX.—Giving the results of feeding G. palpdis on blood through membranes other than freshly-removed
skin
No.
Sex
Membrane
Nature of fluid
Result
Remarks
1
a
Rabbit skin removed
24 hours previously
and partially
decomposed
Defibrinated goat blood
Complete distension
Immediately
2
9
11
»
»»
3
9
Partial distension ...
After 15 minutes
4
a
Ox bladder.
Complete distension
Immediately
5
9
n
»»
11
6
a
Partial distension ...
After 15 minutes
7
a
»>
Slight distension
8
a
Peritoneal
membrane, ox
Partial distension ...
»»
9
9
»
»
Slight distension
»>
10
a
»»
»*
11
9
Thin rubber sheeting
>1
n
12
a
n
Complete distension
!»
*3
9
»>
No visible distension
*4
a
„
»>
*5
a
it
„
16
9
n
is not that the flies require the fluid to be at a positive pressure before
they can imbibe; it is possibly a mechanical difficulty, the fly being
unable to take up fluid unless its proboscis is buried in the membrane
or tissue. The experiments show, moreover, that even when the
proboscis is inserted into a membrane, it does not necessarily follow
that the fluid enclosed will be imbibed to any considerable extent.
The results obtained prove most definitely that Glossina palpalis
exhibits a preference for certain fluids, especially blood, red cells and
haemoglobin. The physical character of the fluid does not, within
limits, appear to matter. Defibrinated blood, a 50 per cent,
suspension of red cells in saline, and a solution obtained by laking
two parts of washed red cells with three of distilled water, are taken
378
up with equal readiness, whilst plasma and normal salt solution are
only imbibed in small quantities as a rule.
Turning to the question whether Glossina takes up vegetable
juices in nature, we may state at once that we have not had the
opportunity of deciding the point absolutely. Glossina of both
sexes were frequently seen to plunge their proboscis into bananas,
oranges and mangoes which had had their skin removed, and also
into the skin itself. The insertion of the proboscis was repeated again
and again, and the organ often remained buried in the fruit for a
minute or more; the whole procedure certainly gave one the
impression that the flies were endeavouring to obtain food from the
fruit. Furthermore, we noticed that Glossina palpalis will pierce
leaves placed on the surface of a fluid with the object apparently of
imbibing the fluid below.
Unfortunately, we had not time to extend our observations on
this subject, but in view of the results already detailed, and of the
observations of Carpenter who found vegetable tissue in a proportion
of wild G . palpalis dissected by him, we consider that it is highly
probable that tsetse do take up food of vegetable nature. The
frequency with which a hungry fly will insert its proboscis, or at
least attempt to do so, into practically any object presented—they
can even be seen trying to pierce the glass vessel in which they are
kept when no skin is in immediate contact with the glass—suggests
that, in the absence of blood, they are on the look out for other food,
for there is every reason to believe that Glossina recognises when it
is in the presence of the skin of a living animal.
Assuming Glossina palpalis cannot reproduce itself or live for
any long period in the absence of blood, it is possible that food of
a vegetable nature may suffice to enable it to exist in a locality which
for a certain period in the year is denuded of vertebrates.
CONCLUSIONS
1. About eight per cent, of the wild G. palpalis in this district
contain recognisable red blood cells—seven per cent, of mammalian
origin and one per cent, nucleated red cells of unknown origin.
2 . Seventy-two hours after G. palpalis had completely distended
itself on rat’s blood recognisable red cells could no longer be found
379
in its intestine; after being fed on a fowl nucleated red blood cells
could be recognised in 40 per cent, of cases at the end of a similar
period. The flies were kept at a temperature of 8o°-86° F.
3. Neither shed blood nor other fluid which is exposed (not
covered by a membrane) can be imbibed by G. palpalis.
4. G. palpalis can take up through a membrane of fresh skin
not only blood and various dilutions of it with normal saline, but
also suspensions of red blood cells in normal saline, and solutions
of haemoglobin (both freshly made from red blood cells, and the
dried crystalline preparation of commerce) in distilled water.
5. Fluids other than blood such as solutions of sugar, sodium
chloride, and glycerine, in water containing a small quantity of a
dye (methylene blue, neutral red or fuchsin) are also taken up
through a membrane of fresh skin by G. palpalis, but not so quickly
or so readily as is blood.
6. G. palpalis exhibits a definite selective taste for the various
fluids presented to it under the membrane; blood, red cells, and
haemoglobin solution being much preferred. The attractive element
in the blood is the fraction of the red cells soluble in water, probably
haemoglobin.
7. G. palpalis which had been starved for a day or two can often
be seen to insert the proboscis repeatedly into oranges, bananas or
other fruits which may be offered them.
8. We are of opinion that G. palpalis in nature may under
certain conditions take up fluid other than blood.
380
REFERENCES
Carpenter, G. D. H. (1912). Progress Report on Investigations into the Bionomics of
Glossina palpalis , July 27, 1910, to August 5, 1911. Report of the Sleeping Sickness
Commission of tbe Royal Society , No. XII, pp. 79-111.
- (1913). Second Report on the Bionomics of Glossina fuscipes {palpalis ) of Uganda.
Reports of tbe Sleeping Sickness Commission of tbe Royal Society , No. XIV, pp. 1-37.
Decent (1909). Sleeping Sickness Bulletin , Vol. I, p. 471.
Lloyd, Ll. (1912). Notes on Glossina morsitans in Northern Rhodesia. Bull. Ent. Res.,
Vol. Ill, pp. 95-96.
- C 1 913)- Glossina morsitans in the laboratory. Ann. *Irop. Med. & Parasit ., Vol. VII,
pp. 285-292.
Maugham (1911). Sleeping Sickness Bulletin , Vol. Ill, p. 271.
Rodhain, J., Pons, C., Vandenbranden, J., and Bequaekt, J. (1912). Contribution au
Mecanitme de la Transmission des Trypanosomes par les Glossines. Arcb. f. Scbiffs- u.
Trop. Hyg.y B. 16, S. 732-739.
Stuhlmann, F. (1907). Beit rage zur Kenntnis der Tsetsefliege. Arb. a. d. Kaiserlicben
Gesundbeitsamte , B. 26, S. 301-383.
Taute, M. (1912). Expcrimentelle Studien iiber die Beziehungen der Glossina morsitans
zur SchUfkrankheit. Zeitscbr. f. Hygiene , B. 72, S. 316-320.
THE CULTIVATION OF THE LEPROSY
BACILLUS
HENRY FRASER, M.D. Aberd.
DIRECTOR, INSTITUTE FOR MEDICAL RESEARCH, FEDERATED MALAY STATES
AND
W. FLETCHER, M.D. Cantab.
PATHOLOGIST, INSTITUTE FOR MEDICAL RESEARCH, FEDERATED MALAY STATES,
KUALA LUMPUR
(Received for publication 16 ]une y 1915)
In an article entitled ‘ Leprosy: a perspective of the results of
experimental study of the disease/ by Bayon, and published in the
Annals of Tropical Medicine , Vol. IX, the statement is made on
page 29 that ‘ Fraser and Fletcher discarded all diphtheroids because
of their ubiquity/ It is unfortunate that he has quoted our work
incorrectly. In the Lancet , Vol II, 1913, we published an article
entitled ‘The Bacillus leprae : has it been cultivated?* and on
page 920, under the head of contaminating micro-organisms, we
state that * In common with other workers we have isolated
diphtheroid organisms, but these are ubiquitous and demand no
special consideration.* At the time that article was written, our
investigations had led us to form the opinion that the diphtheroids
were contaminators, and had no genetic relationship with the leprosy
bacillus; further investigations carried out during the past two years
have confirmed that opinion.
If we had confined our observations to the results of one or two
experiments, as has been done by so many workers, we might,
perhaps, like Bayon, have formed the opinion that the diphtheroids
were related to the leprosy bacillus, but to do so we should have had
to reconcile or, since that was impossible, to ignore some remarkable
discrepancies.
It seems a comparatively simple operation to reflect the skin from
a leprous nodule and to excise a portion of the subcutaneous tissue
free from contamination, but only those who have performed the
operation a sufficient number of times can be aware of the pitfalls.
Cultures of diphtheroids were obtained only in our early experi¬
ments ; in these a nodule of tissue rich in leprosy bacilli was excised,
and from twenty to thirty tubes of culture media were inoculated
382
with portions of it. On one tube, perhaps two tubes, a culture of a
diphtheroid would be obtained. As each tube was inoculated with
an enormous number of leprosy bacilli from the same nodule,- it is
impossible to believe that only in one or two parts of that nodule
was there an acid-fast bacillus capable of proliferation under
saprophytic conditions into a diphtheroid. If every tube inoculated
with portions of the nodule had developed a culture of a diphtheroid
the opinion formed might have been quite different, but the sporadic
occurrence of a diphtheroid is quite in accordance with our results
obtained in the isolation of diphtheroids from other parts of the
body, and we believed the correct interpretation of these experiments
to be that the nodule was contaminated, or had become contaminated
from the skin, with one or two diphtheroid bacilli which proliferated
on the culture media.
In that belief we extended our investigations and improved our
technique, avoiding the use of all disinfectants for the skin. We
were then able to excise leprous nodules free from contamination,
and from which a culture of no organism could be obtained on any
medium.
We have now excised material from fifty-two non-ulcerating
nodular cases of leprosy, a number far in excess of that recorded by
any other worker, and have employed the media of every claimant
to success, but have not substantiated the claims of any of them.
Our work has been carried out over a period of three and a half
years The only conclusions, which that work permits, are that the
leprosy bacillus has not been cultivated and that the diphtheroids
and other organisms are merely contaminators.
Elsewhere we (1914) have dealt with Kedrowsky's culture of an
acid-fast bacillus which Bayon claims to be the leprosy bacillus.
Our experiments have led us to conclude that there is no evidence
that the acid-fast bacillus of Kedrowsky is the leprosy bacillus.
REFERENCES
Bayon, H. (1915). Leprosy: a perspective of the results of experimental study of the
disease. Annals Trop. Med. & Parasitol ., IX, pp. 1-90.
Fxasxr, H., and Fletchki, W. (1913). The Bacillus leprae: has it been cultivated?
Lancet , Sept. 27, 1913, pp. 918-921.
- - ( I 9 , 4)- Fourteenth Annual Report of the Institute for Medical
Research, Kuala Lumpur, Federated Malay States.
383
THE RESERVOIR OF THE HUMAN
TRYPANOSOME IN SIERRA LEONE
[Being the Third. Report of the Thirty-second Expedition of the
Liverpool School of Tropical Medicine, 1914-1915.]
BY
WARRINGTON YORKE
AND
B. BLACKLOCK
(Received for publication 24 June , 1915)
No time was spent in examining the native population for the
incidence of trypanosomiasis, but two cases of sleeping sickness—
a woman and her child—were brought to our notice by Drs. Young
and Butler of the West African Medical Service. These cases which
were extremely chronic were discovered accidentally, one or two
trypanosomes being seen during the examination of the blood for
malarial parasites. The patients lived in Lumley, a native village
situated at the junction of the Cape Lighthouse Peninsula with the
mainland, about a mile and a half from Hill Station and four miles
from Freetown. Trypanosomes were first seen in the mother
(Catherine Macauley) about the beginning of 1914. She received a
single treatment with salvarsan, and subsequently returned to
Lumley. When seen by us on December 7th, 1914, she was in good
condition. According to Dr. Young, no definite change had
occurred during the year. The only symptoms were headache and
a feeling of weakness. She had one or two slightly enlarged glands
in the posterior triangles. Several careful examinations of the
peripheral blood and gland juice failed to reveal trypanosomes. On
the day she was first seen by us (December 7th) two rats were
inoculated with blood and one with gland juice. After 28 days one
of the two rats inoculated with the blood was found to be infected
with trypanosomes. The other two rats were kept under observa¬
tion until they died, on the 59th and 67th days respectively;
trypanosomes were never found in them. There was no change in
the condition of the patient when she was last seen by us on
February 24th, 1915. On December 16th, 1914, she brought her
son, aged 8, to the hospital suffering with fever. On examination
of a stained preparation of his blood by Dr. Butler, malarial
parasites were found and also a single trypanosome. On
December 19th the boy was seen by us. He complained of pain
over the spleen, which was enlarged, reaching to within one inch of
the umbilicus. There were a few shotty glands in the neck, axillae
and groins. Malaria parasites were still present in the blood, but
no trypanosomes were seen. Two rats were inoculated with 1 c.c.
of the peripheral blood. Neither of these animals became infected;
one died on the 52nd day afterwards, and the other was still alive
on the 90th day when we left the Colony. The blood of this boy
was examined by us on several other occasions, but trypanosomes
were not seen.
The history of these two cases shows that sleeping sickness in
Sierra Leone is exceedingly chronic, and very difficult to recognise.
Trypanosomes can be found only occasionally in the peripheral
blood; gland puncture of the mother was negative, whilst the boy
had no glands which were puncturable.
The question whether there is any reservoir of the human
trypanosome in Sierra Leone other than man is of importance, and
is one which we decided to investigate. By the term Reservoir we
mean the source from which Glossina derives its infection.
Kinghom and Yorke (1912) have shown that the chief reservoir of
the human trypanosome of South Central Africa (T. rhodesiense )
is the antelope and not man. The antelope differs from man in
being tolerant of the infection; unlike man, in whom the disease
runs an acute course, it is able to harbour T. rhodesiense in its
blood for long periods without exhibiting signs of the disease. Both
for this reason and because the antelope exists in enormous numbers
in many parts of South Central Africa, coinciding in its distribution
with the ubiquitous G. morsitans —the fly responsible for the
transmission of sleeping sickness in this region—it constitutes a
much greater and more reliable reservoir of the human trypanosome
than does man. In Sierra Leone, however, we have to deal with
quite a different state of affairs; sleeping sickness in this part of
Africa is an extremely chronic infection. In striking contrast to
3»S
cases infected with T. rhodesiense , those infected with the human
trypanosome of the West Coast may live for many months, or even
years, without exhibiting any serious indication of the disease. For
this reason, therefore, man in Sierra Leone is a more constant and
dependable reservoir of the virus of sleeping sickness than he is in
Rhodesia or Nyasaland. Moreover, in the Colony of Sierra Leone,
and in most portions of the Protectorate, large game is compara¬
tively rare. Again, the most prevalent tsetse-fly in Sierra Leone
is G. palpalis ; owing to its predilection for water-courses its distri¬
bution is limited much more closely to the haunts of man than is
that of the ubiquitous G. morsitans. In Sierra Leone, therefore, we
must conclude that of large game and man, the latter constitutes by
far the more important reservoir of the human trypanosome.
There is, however, another possible reservoir of the human
trypanosome which must not escape attention, that is domestic
stock. The Sleeping Sickness Commission of the Royal Society
(1910) examined seventeen cattle in Uganda, and found one cow
infected with T. gambiense. This element does not exist in
Rhodesia or Nyasaland, as quite apart from T. rhodesiense cattle
are unable to thrive to any extent in the presence of large numbers
of G. morsitans. Although cattle are not bred in most portions of
Sierra Leone, yet they are imported from French Senegal in large
numbers, and gradually find their way down to Freetown for
slaughter.
Both in the Protectorate and in the Colony the blood of a
number of domestic animals—143 animals, 7 goats, 7 sheep and
10 dogs—was examined. As will be seen in a subsequent paper,
the trypanosomes most commonly found were T. vivax and
T. congolense. One ox, however, was found to be infected with a
trypanosome which we are unable to distinguish from that infecting
man. The ox in question was one of a herd of 90 Government
cattle in a Warri at Batkanu. All the animals in this herd had
originally come from French territory, but they had been in the
Protectorate for various periods before being bought by the Govern¬
ment. The animal in which this trypanosome was found appeared
to be in perfect health; direct examination of its peripheral blood
was negative. The trypanosome was discovered fortuitously owing
to the fact that this ox happened to be one of nine chosen at random
from the herd for the purpose of having their blood inoculated into
rats. Trypanosomes were seen in the peripheral blood of the rat
fourteen days after inoculation. The parasite was a polymorphic
trypanosome, it did not exhibit posterior nuclear forms, and was
indistinguishable morphologically from the trypanosome isolated
from the human case of trypanosomiasis.
During the last six months the strain has been maintained by
passage through rats. The course of the infection in these animals
is exceedingly chronic, and certain rats failed to become infected.
For purpose of comparison, the result of inoculation of rats and
guinea-pigs with this strain and with that obtained from man are
given in tabular form.
There is a striking similarity in the pathogenicity of the two
strains in rats. The virulence of both is but slight, and is in marked
contrast to that of the other common polymorphic trypanosomes—
T. pecaudi or T. rhodesiense vel ugandae (T. brucei , Uganda). On
consulting the results of inoculation of rats with T. gambiense direct
from man (Yorke, 1910) one cannot fail to observe their striking
similarity to those set forth in the above table. Macfie (1914)
records that he failed to infect rats with the Nigerian strain of the
human trypanosome (T. niget tense). A guinea-pig infected with
T. nigeriense was, however, sent by Macfie to us at Runcorn, and
from it we succeeded without much difficulty in infecting a number
of rats. The course of the infection in these animals was
exceedingly chronic often ending in recovery; some failed to become
infected.
We are of opinion, therefore, that this polymorphic trypanosome
from the ox is identical with that infecting man in West Africa,
in other words, that it is T. gambiense .
The discovery of the human trypanosome in the ox is important.
It shows that domestic stock may serve as a reservoir of
T. gambiense. Before we can form any opinion as to the relative
importance of this reservoir we must have information on the
following two points : —
(1) Do the animals harbour the parasite in their blood for long
periods without exhibiting signs of disease, or does the infection
run an acute course?
(2) What percentage of the animals is infected ?
3«7
Table I.—Giving results of inoculation of rats with the human strain of T. gambiense
No. of Animal
Animal from which
inoculated
Incubation
in
days
Length
of life
in days
Remarks
Rat la .
Patient’s blood
28
■56
» ib .
33 j»
—
Did not become infected
» io* .
Rat la .
8
88
» iob .
» « .
13
—
Alive on 171st day.
Trypanosomes last seen
on 50th day
» 37 * .
33 * oa .
4
29
» 37 ^ .
>» ioa .
—
—
Did not become infected
33 5 Ia .
n loa .
—
-
33 >»
>1 5*b .
5j i° a .
—
—
» i»
Table II.—Giving results of inoculation of rats and guinea-pigs with the ox strain
of T. gambieiue
No. of Animal
Animal from which
inoculated
Incubation
in
in days
Length
of life
in days
Remarks
Rat 24 .
Ox VIII .
14
40
» 3 i .
Rat 24 .
7
—
Alive on 160th day.
Trypanosomes last seen
on 24th day
» 47 a .
» M .
10
39
» 47 b .
3) H .
10
—
Alive on 139th day.
Trypanosomes last seen
on 67th day
33 49 ® .
3, 3 » .
—
-
Did not become infected
t> 49 b .
33 3 i .
—
—
13 11
33 50 .
33 47 b .
10
63
Guinea-pig 52
33 5 ° .
—
—
Did not become infected
Rat 53* .
33 50 .
18
-
Alive on 46th day
». 53 b .
33 5 ° .
18
—
33 33
Guinea-pig 54
33 53 a .
—
—
Did not become infected
55 .
33 53 a .
*3
—
Alive on 31st day
388
As regards the first point, the ox from which the trypanosome
was obtained appeared in perfect health, and a report received four
months later states that there is still no indication of disease.
Regarding the second point, we have as yet no evidence to show
what proportion of the cattle in Sierra Leone are infected with
T. gambiense. If, as appears probable, cattle prove to be tolerant
of this parasite and can harbour it in their blood for long periods
without detriment to health, it will be no easy matter to determine
how many animals are infected. As already mentioned, the
parasite was discovered largely by chance; although a direct
examination of the peripheral blood of each of the 90 animals in
the Warri was made, subinoculation into rats was done in nine
instances only. Furthermore, it must be borne in mind that rats
not infrequently fail to become infected after inoculation of blood
containing T. gambiense. It will, therefore, probably be at least as
difficult to determine the percentage of cattle infected as it is to
ascertain the number of infected human beings.
Before leaving the subject, it is interesting to refer to the
observations recorded by Macfie. This investigator holds that in
the Eket district man does not constitute a reservoir of sleeping
sickness, and that the reservoir is still unknown. The evidence on
which he bases this assumption is, in our opinion, unconvincing.
He writes 4 The extreme rarity or complete absence of trypanosomes
from the peripheral blood of all the human cases examined, the
rarity of the parasites even in the gland juice, and the difficulty
experienced in infecting animals by inoculation, suggest that it
must be a very exceptional occurrence for a tsetse-fly to become
infected by feeding on these cases.It seems, therefore, that
the human infections must be dependent on some other cycle of
development, including an insect and some so far unidentified
animal host, the reservoir of the disease. The ordinary development
of the trypanosome may take place in these two hosts. The insect,
infected from the animal host may, however, be capable of infecting
human beings; but the disease may be so modified in them that they
are incapable of handing on the infection any further/ This
argument appears fallacious. In view of the fact that trypanosomes
were not found in the peripheral blood they were doubtless scanty,
but it does not by any means follow that they were absent. Nor
389
does the failure to infect animals by inoculation of blood materially
assist the argument, since Macfie himself found what has frequently
been pointed out by previous workers that rats and guinea-pigs
often fail to become infected after the injection of blood known to
contain T. gambiense. Macfie records that all his attempts to
infect rats with blood containing the trypanosomes were unsuccessful;
further, of seven guinea-pigs inoculated with gland juice actually
containing trypanosomes only one became infected.
This comparative insusceptibility of rats and guinea-pigs to
human trypanosomes of the West Coast shows that we cannot safely
regard a negative result obtained by inoculation as evidence that
trypanosomes were not present in the. blood inoculated. It is,
therefore, inadmissible to argue, as does Macfie, from the negative
results of microscopic examination of the blood and of inoculation
of guinea-pigs and rats that G. falpalis cannot become infected from
human beings. In 5 c.c. of the blood of one of these patients there
may be sufficient trypanosomes to infect many flies, and yet
insufficient to infect a particular guinea-pig or rat.
Briefly, Macfie’s view appears to be this. Man is not a reservoir
of the virus of sleeping sickness because Glossina cannot become
infected from him. He postulates an unknown vertebrate reservoir
of the virus and an insect host, in which the ordinary development
of trypanosomes takes place. The insect may infect man but cannot
become infected from him, but only from the hypothetical vertebrate
reservoir.
For the reasons given above, we consider that Macfie has failed
to show that Glossina cannot become infected from man. In our
opinion, there is no evidence which would lead us to believe that man
does not constitute a most important reservoir of the human
trypanosome of West Africa. Nevertheless, the fact that the human
trypanosome has been discovered in an ox indicates that domestic
stock also forms a reservoir, the extent of which we are at present
unable to estimate, but the existence of which must be recognised
when prophylactic measures are contemplated.
CONCLUSIONS
1. The human trypanosome, T. gambiense , has been found in
an ox in Sierra Leone.
390
2. It is impossible at present to form any conclusion of the
extent to which domestic stock may harbour this parasite. In all
probability, it will prove to be at least as difficult to recognise the
infection in cattle as it is in man.
3. The existence in domestic stock of a potential reservoir of
T. gambiense will have to be taken into account when prophylactic
measures are contemplated.
REFERENCES
Bruce, Hamerton, Bateman, Mackie and Lady Bruce (1910). Experiments to ascertain if
cattle act as a reservoir of the virus of Sleeping Sickness ( Trypanosoma gambiense). Free.
Roy. Soc. B. VoL 82, pp. 480-484.
Kinghorn, A. and Yorke, W. (1912). On the Transmission of Human Trypanosomes by
Glossina mortitans, Westw.; and on the Occurrence of Human Trypanosomes in Game.
Annals Trap. Med. and Parasitol ., VI, pp. 1-24.
Macfie, J. W. S. (1914). Sleeping Sickness in the Eket District of Nigeria. Annals Trap.
Med. and Parasitol ., VIII, pp. 379-438, with 5 plates. 1 map.
Yorke, W. (1910). On the Pathogenicity of a Trypanosome (T. rbodesiense , Stephens and
Fantham) from a case of Sleeping Sickness contracted in Rhodesia. Annals Trap. Med •
and Parasitol ., IV, pp. 351-368.
39*
SPIROCHAETA BRONCHIALIS ,
Castellani, 1907, TOGETHER WITH
REMARKS ON THE SPIROCHAETES OF
THE HUMAN MOUTH
BY
H. B. FANTHAM, M.A. Cantab., D.Sc. Lond.
LECTURER ON PARASITOLOGY, LIVERPOOL SCHOOL OP TROPICAL MEDICINE.
[Being the First Report of the Thirty-first Expedition of the
Liverpool School of Tropical Medicine , to Khartoum , 1913]
{Received for publication 1 ]une> 1915)
Plate XXXIV
CONTENTS
I. Introduction. 391
II. Material and Methods . 392
III. Movements . 394
IV. General Morphology . 395
V. Morphological Variation in Spirocbaeta broncbialu . 396
VI. The Granule Phase or Spirocbaeta broncbialu . 399
VII. Some Further Remarks on the Granule Phase or Spirochaetes. 401
VIII. The Spirochaetes or the Human Mouth . 402
IX. The Spirochaetes or the Human Mouth contrasted with S. broncbialu ... 404
X. Mode or Intection. 404
XI. General Remarks . 405
XII. Summary and Conclusions. 408
Reverences . 409
Explanation or Plate . 412
I. INTRODUCTION
During the summer of 1913, I had the good fortune to work in
the Wellcome Tropical Research Laboratories, Khartoum, having
been sent on expedition by the Liverpool School of Tropical
Medicine. While there, thanks to the kindness of the Government
of the Anglo-Egyptian Sudan and of the Director, Dr. A. J.
Chalmers, and Staff of the Wellcome Laboratories, I was able to
39 2
undertake researches on human and avian spirochaetes, directing
especial attention to bronchial spirochaetosis in man. In this
memoir an account is given of some of my researches on the causal
agent of the latter disease.
II. MATERIAL AND METHODS
The material was chiefly obtained from cases of human bronchial
spirochaetosis occurring in Khartoum, Omdurman and Kodok.
Many of the cases were chronic. They included officers’ servants,
warders, native policemen, soldiers and household servants. A few
cases among Europeans suffering from bronchitic symptoms were
studied, and some of them were found to be fresh cases of undoubted
spirochaetosis. Among the latter were two cases of almost experi¬
mental infection. The total number of cases studied was twenty.
I was fortunate in being able to obtain material from nearly all
the cases reported by Drs. Chalmers and O’Farrell (1913). I was
also able to examine preparations sent by Dr. J. A. Taylor from
Entebbe, Uganda, to whom my best thanks are due. In fact,
Dr. Taylor sent me his first preparations for my examination and
opinion at the end of May, 1913, just before I left England for the
Sudan. Spirochaeta bronchialis was also studied in monkeys
experimentally infected in Khartoum.
It is with much pleasure that I acknowledge the kindness of
Dr. Chalmers and Captain O’Farrell, R.A.M.C.; Major Forrest,
R.A.M.C.; Major Carroll, R.A.M.C.; Dr. Christopherson;
Dr. Atkey; Dr. Crispin; Captain Stirling, R.A.M.C., and
Captain Buist, R.A.M.C., in providing me with material for research.
I am especially indebted to the gentlemen first-named for much help
while in the Sudan.
The subject is one of great difficulty, and because of this
difficulty and the pressure of other work the publication of this paper
has been delayed. Also, even now, it is not so fully illustrated
as I should have wished. Many more drawings (totalling over 500)
have been prepared, but owing to the war these cannot be reproduced
without further delay. Hence a selection of what appear to be
types of the spirochaetes only are shown on the accompanying
plate (XXXIV), the reproduction of which by half-tone process has
left something to be desired.
393
Also, owing to the lack of precise knowledge of spirochaetes
found in the human mouth and respiratory passages, attention has
been paid to the comparative morphology of these organisms. I
have compared the spirochaetes of the mouth found in natives of
the Sudan with those found in similar situations in people living in
England, especially in cases of pyorrhea alveolaris and of ‘ gum¬
boils,* obtained in Liverpool and in Cambridge.
In the Sudan the procedure followed for obtaining material was
briefly as follows:—The teeth were examined for caries, as were
also the buccal, tonsillary and pharyngeal secretions, with a view
to finding spirochaetes. The patient washed his mouth with clean
water, and samples of spirochaetes in mouth sputum might also be
thus obtained. The throat was then gargled, and usually samples
of throat sputum were examined. After this careful washing of the
mouth and gargling of the throat, expectoration from the deeper
bronchial regions was collected in a sterile petri-dish, and in this
Spirochaeta bronchialis occurred. The sputum thus collected was
examined by dark-ground illumination, and films were made with
the aid of a sterile platinum loop for immediate fixation and
staining.
Various methods of fixing and staining were tried. Osmic
vapour followed by absolute alcohol, Schaudinn's fluid (sublimate-
alcohol-acetic) and Bouin*s fluid (picro-formol-alcohol-acetic) were
used for fixing wet preparations; Bouin*s fluid was found most
useful. The solutions of Giemsa and Leishman, carbol thionin and
gentian violet, were tried, as well as various haematoxylins, such as
those of Delafield, Ehrlich and Bohmer. In the hot dry atmosphere
of Khartoum, freshly prepared Bohmer’s haematoxylin was found
to be most useful, and less complicated than iron-haematoxylin,
though this Was also used. Staining overnight with dilute
solutions was found to be more effective in the case of this
fragile spirochaete than quick staining with stronger solutions.
Unfortunately, gentian violet, so useful for spirochaetes in general,
was found in Khartoum to fade quickly, but was very useful when
studying mouth spirochaetes in England. Naturally, in the fixation
of a spirally wound, flexible organism, some flattening and
straightening of the coils is inevitable, the fixed forms thus being
less graceful structures than the living ones.
394
III. MOVEMENTS
In fresh sputum 5 . brottchialis moves very quickly. Its move¬
ment, like that of other spirochaetes, is difficult to analyse. The
organism moves forward while turning on its long axis. However,
as I have previously described (1907, 1908), such movements may
be divided into two components, namely, an undulatory flexion of
the body mainly for progression, and a corkscrew or helicoid move¬
ment of the body as a whole, due to the winding of the membrane
or crista. This latter organella is very narrow and thin, and is a
lateral outgrowth of the periplast. It is only seen with difficulty in
S. brotichialis , and not in some specimens, as it is sometimes so
contracted as to appear to be absent. It does not markedly
undulate, and should not be called an undulating membrane in
spirochaetes, as I pointed out in 1907-8, for it is not directly
comparable with the undulating membrane of a trypanosome. It
can be seen in some cases in life by the use of the paraboloid
condenser. In stained specimens it sometimes appears as a curved
line lying along the body when the organism has been Axed during
rapid movement (PI. XXXIV, Ags. 2, 22), or as a slightly wavy
lateral outgrowth when the organism was killed while moving slowly,
the membrane being somewhat relaxed (Ags. 10, 16, 36).
The undulatory flexion is responsible for the rippling motion of
spirochaetes and the waves seen passing down the body of these
organisms.
Slowly moving spirochaetes have few undulations of relatively
large amplitudes along their bodies, while quickly moving forms
have more numerous small undulations (Ags. 19, 23, 28), characters
which I also described in 1907-8. Hence the number of coils or
waves in a spirochaete is more an index of its rate of motion—as well
as of its thickness,which is also a factor—than one of differentiation
between various forms. Thus, thin forms are more easily thrown
into waves than thick forms, and so the number of waves in a
spirochaete cannot be used as a mark of species differentiation as
has been attempted by some workers in the past. In spirochaetes
exhibiting polymorphism, thin forms represent young individuals,
while thick ones are adult forms, as was indicated by me in 1909.
Jerking movements may occur, but not frequently. Spirochaetes
are isopolar, and can easily and very quickly return on their own
path. Other remarkable appearances are produced by rapid move-
395
ments in a relatively limited area. Thus, the spirochaete may curve
each of its ends inwards in a watchspring-like manner and after a
time straighten itself, and then re-form a watchspring coil at either
end in the opposite direction. Waves continue to pass along the
organism during the movements of the body as a whole. At
other times, though more rarely, the organisms intercoil their ends
and seem almost to tie themselves in knots, only to extend a few
seconds later and swim away in a different direction. The subject
of the movements of spirochaetes was fully discussed by me in June
and August, 1907, and January, 1908, when dealing with
S. balbianii and S. anodontae , and the same remarks apply to
S. bronchialis and the spirochaetes of the mouth. There was little
evidence of division seen in life.
IV. GENERAL MORPHOLOGY
The morphology of spirochaetes is a subject of great difficulty
and of no little controversy. In this paper I shall, without prejudice,
adopt the generic name Spirochaeta, as I cannot at present accept the
unconfirmed researches of Zuelzer (1911) on that debatable organism
known as S. plicatilis. As a result of Zuelzer’s researches some
workers have proposed to create a number of genera such as
Cristispira (for the forms found in molluscs), Spiroschaudinnia or
Borrelia (for blood-inhabiting forms), Treponema , and the like, but
without any clearly and generally accepted generic differences. The
divergent descriptions of S. plicatilis given by different workers may
be clearly seen and contrasted from the account given in Bosanquet's
book on Spirochaetes. The so-called axial fibre of Zuelzer is
acknowledged to be homologous with the membrane or crista of
molluscan spirochaetes. Subsequent researches may lead to a
solution of these difficulties.
Spirochaeta bronchialis is a delicate organism which frequently
stains with difficulty. It exhibits morphological variation (figs. 1-39),
due to growth and division. There are long forms (figs. 37-39)
and short forms (figs. 1-6); also thick ones (figs. 7, 38) and thin
ones (figs. 28, 29, 33), and those of intermediate length (figs. 7-33)
and breadth (figs. 25, 34, 35). No detailed account of the morphology
of the parasite has yet been published.
The cytoplasm of S. bronchialis is almost homogeneous and
39 $
shows no vacuoles. In spite of the minuteness and fragility of the
organism, chromatin bars can be seen at intervals along the parasite
in some of the specimens (figs. 14, 24, 29, 36-39). In stained
preparations the observance of these minute bands or rodlets is some¬
times aided by the use of stereoscopic eyepieces. When examined
fresh these chromatin granules appear as retractile spots, as
viewed under dark-ground illumination using a paraboloid
condenser. On the other hand, many spirochaetes only show the
chromatin dots or granules with difficulty at certain stages of their
life-cycle, and so may appear more or less homogeneous (figs. 19,
28, 34)-
The presence of chromatin bars or rodlets at short intervals
along the spirochaete in a stained preparation (figs. 24, 29, 32) gives
rise to a so-called alveolar or chambered appearance, which has been
much emphasised by Gross (1910) and those who have followed him.
There is little or nothing new in Gross’s observations, except the
terms used to express his interpretations. However, his views have
been so dogmatically and even polemically asserted that work
previous to that of Gross has tended to be overlooked, although it
was performed in many cases with due regard to careful cytological
technique. The accentuation of differences of interpretation is
merely indicative of partisanship, and does not conduce to the
progress of knowledge.
The ends of S. bronchialis are tapering. Observed separately
they may appear to be pointed (figs. 26, 33) or somewhat rounded
(fig. 7) in stained preparations. Sometimes one end appears rather
more tapering than the other (figs. 31, 39), a feature that is
explained by extending attenuation at a previous division. So-called
flagella are also thus explicable. These variations in the character
of the ends of S. bronchialis also occur in other spirochaetes, such as
S. balbianii , and were described by me at some length in 1909.
V. MORPHOLOGICAL VARIATION IN SPIROCHAETA BRONCHIALIS
Spirochaeta bronchialis exhibits considerable polymorphism,
resulting from the processes of growth and division. This range of
variation is responsible probably for the varying dimensions of the
organism given by different workers. The difference of size
397
exhibited by S. bronchialis can only be realised by examination and
measurement of a large number of specimens derived from a series
of cases. Measurements of spirochaetes from one or two cases only
may be very fallacious, since the organisms observed may be all
practically at the same stage of development.
By measuring a moderately large number (300) of bronchial spiro¬
chaetes, the discrepancies in length may be explained; I find that the
length ranges from 5 p to 27 p. The size of a number of them
centres around 15 p, while many of the others are about 8 p
long. Castellani and Chalmers (1913) in their 'Manual of Tropical
Medicine/ state on p. 1283 that one form of S. bronchialis is from
15 P to 39 At, while on p. 402 the commonest form is said to be
7 p to 15 p long. Thus my measurements give a lower limit for both
minimum and maximum dimensions as stated by them, while a
number of the spirochaetes that I have measured are about 15 p,
Macfie (1915) states that his smallest form was 6 p long, his
largest 13 p , and that the average length of his specimens was 8 p to
9 p . When examining my series of slides, I found one—taken from
a chronic case on a particular day—on which the spirochaetes seemed
remarkably uniform. A number was drawn with the aid of a camera
lucida and measured (figs. 7, 12, see also 13). It was found that
almost everyone of them was from 8 p to 9 p long. Their measure¬
ments coincided almost exactly with those given by Macfie, and
had this slide been the only one, erroneous conclusions would have
resulted. Measurements of the spirochaetes from the same case on
the days preceding and on subsequent dates showed that the range
of length of 5 . bronchialis was considerably greater than was shown
on that one occasion. As Macfie's dimensions were derived from
two cases only, it was likely that the majority of the spirochaetes
were at the same stage of development, and thus had attained about
the same size. Many bronchial spirochaetes from Khartoum were
either from 14 p to 16 p long, or 7 p to 9 p long. Definite
evidence of transverse division in members of the former group
has been obtained, and it is very likely that Macfie’s forms
measuring 8/1 to 9 m so originated. Chamberlain (191 1) having
found bronchial spirochaetes in two typhoid patients in the
Philippines, states that their average length is 15 p. Some of
the spirochaetes that I examined in the Sudan measured about
398
15 ft long, but much depended on the age of the parasites and on
the condition of the patient. For instance, the predominant type
of S. bronchialis found in several chronic cases was an organism with
two tapering ends, one sometimes slightly more rounded than the
other, with a length of 13 ft to 16 ft (figs. 34, 35).
Dr. Taylor kindly sent me some slides of S. bronchialis from
Uganda. I have drawn and measured spirochaetes from each of the
slides he sent me, and can state that not only can I recognise the
four types mentioned by him, but that the dimensions which I have
obtained from his sketches agree in the main with mine (figs. 40-45).
The range of length obtained from my measurements is from 5*5/1
to 19 5 ft. Dr. Taylor, in his Report for 1913, differentiated four
groups of spirochaetes, which I have determined from his figures to
measure from 6 fit to 9 fit, 6 fit to 8 fit, 15 fit to 18 ft and 9 ft to 11 ft,
respectively. The first three were considered by him to be bronchial
spirochaetes, the last occurred in the mouths of healthy persons.
While the spirochaetes were thus grouped by Dr. Taylor, he recog¬
nised the existence of possible transitional forms and that one group
gradually merged into another.
The length of S. bronchialis is often a factor of its age. Forms
recently developed by the elongation of granules or coccoid bodies,
naturally, are short (fig. 1-6). Small forms also arise by transverse
division of older, longer ones. The nutrient medium in which the
spirochaetes occur also seems to react upon their morphology.
Organisms surrounded by thick stringy mucus may become stouter
than those in more fluid sputum, which offers less resistance to their
passage through it. Spirochaetes found in a denser medium tend
to be both shorter and broader than those in a more fluid one in
many cases, though no general statement can be made regarding this
factor. The degree of relaxation obtaining in the organism when it
was killed is also a factor in producing differences in the appearance
of the coils of the organism, and consequently in its dimensions.
Polymorphism or morphological variation of S. bronchialis occurs
not only with respect to length, but also in connection with breadth
and the character of the ends of the organism. The breadth of the
spirochaetes shows some variation, but it is less marked than the
variations in length. Occasionally a spirochaete broader than the
average is found (fig. 38). The breadth is not easily determined,
but varies from 0*2 ft to 0*6 ft .
399
The ends of the spirochaetes show considerable variation. The
degree of tapering manifested by them depends in part on the motion
of the parent organism during division, as mentioned previously.
Spirochaetes in which both ends taper, but wherein one is somewhat
more pointed than the other, are fairly common (figs. 27, 34, 37, 39,
46). Others show the ends equally pointed (figs. 8, 10, 22, 26); yet
others have relatively rounded ends after some tapering of the body
has occurred (figs. 7, 31, 47, 48). Extreme tapering of one or both
ends of the spirochaete have led to the erroneous interpretation of
flagella in the case of other spirochaetes, such as S. dentium .
Occasionally such an interpretation might be made in specimens of
5 . bronchialis , but examination of many of the organisms, both
living and in stained preparations, confirms the fact that true flagella
do not occur in the organism.
A membrane or 4 crista* is present in S. bronchialis (figs. 10, 16),
but is not always easy of detection. This difficulty of demonstrating
the membrane would, perhaps, cause some workers to assign the
organism to the genus Treponema , in which hitherto a membrane has
not been satisfactorily demonstrated. Such classification, however,
would be fallacious, inasmuch as the coils of a Treponema are said
to be pre-formed while those of a spirochaete are not, but are capable
of variation according to the rapidity or slowness of motion and to
the density of the medium traversed.
VI. THE GRANULE PHASE OF SPIROCHAETA BRONCHIAUS
The formation of coccoid bodies or granules in Spirochaeta
bronchialis has been studied in fresh preparations, using both
ordinary illumination and the paraboloid condenser, as well as in
stained preparations. As the organisms are small, there is some¬
times some slight difficulty in studying the process of formation of
these minute reproductive bodies, though it is relatively easy to
observe in larger, somewhat stouter specimens about 11 n to 16 fi long
and about 0*3 fi broad examined at the right stage of development.
Further, there seems to be a periodicity in the formation of granules
in S. bronchialis , but the exact period has not yet been determined.
The process of formation as observed in life is as follows: —
The cytoplasm at first is very finely granular, in fact, almost
homogeneous. The chromatin bars appear as minute refractile
400
masses. A concentration of some of the cytoplasm occurs around each
chromatin rodlet. These small concentrations gradually become
oval, the outer cytoplasmic layer differentiates as a thin coat, and
ultimately a series of coccoid bodies or granules is formed
(figs. 46-49), lying usually transversely or slightly obliquely within
the periplast sheath. Sometimes the coccoid bodies are set at
liberty by a rupture appearing at one end of the spirochaete (fig. 50),
at other times several ruptures, or disintegration of the sheath can be
observed. A few empty sheaths have been found both in fresh and
in stained preparations. Stained specimens show a series of darker
lozenge-like coccoid bodies alternating with relatively clear, pale
staining areas (figs. 47-49).
The formation of coccoid bodies takes place both in free
spirochaetes and in those which have penetrated the delicate cells
lining the air passages (fig. 52). Sometimes two or more spirochaetes
have been found within a mononuclear cell. They may be in the
ordinary trophic phase, or may be in process of formation of
coccoid bodies. When within a cell the coccoid bodies often
seem to be liberated by the disintegration of the periplast. Groups
of coccoid bodies still retaining the outline of the spirochaete from
which they originated (fig. 52) are of fairly frequent occurrence.
When the coccoid bodies are released by a terminal rupture of the
parent, they tend to form irregular clumps. The elongation of the
granules and the emergence of very small spirochaetes from the
groups of granules (fig. 51) have been observed in life. It is very
probable that there is a definite period in the life of a spirochaete at
which there is a marked differentiation of coccoid bodies. It must
also be borne in mind that coccoid bodies may be present when
spirochaetes as such cannot be detected.
In staining reactions, size and morphology, the coccoid bodies
are different from any bacteria which may be present in the
surrounding medium. They stain less darkly than cocci, and are
smaller (fig. 52). Spirochaetal coccoid bodies have no capsule such
as occurs commonly in bacteria.
4oi
VII. SOME FURTHER REMARKS ON THE GRANULE
STAGE OF SPIROCHAETES
In continuance of the remarks which I made on the subject of the
granule stage of spirochaetes in these Annals (1914), Vol. VIII,
pp. 471-484, I should like to draw attention to the following further
evidence.
1. Noguchi, in his address before the Royal Society of
Medicine, London, on October 20th, 1913, stated that he ‘was able
to demonstrate .... granules in the pure cultures of Treponema
pallidum . This phenomenon, however insignificant it may appear
in itself, was destined to furnish a key to one of the most disputed.
problems of the past fifty years—namely, the problem of so-called
parasyphilis, since it was this very idea that prompted me to
undertake to search for Treponema pallidum in one form or another
in the brains of general paralytics and in the spinal cord from cases
of tabes dorsalis.' And again, ‘ I was led by the observation that
Treponema pallidum sometimes assumes a granular form in cultures
to re-study sections of paretic brains stained for th z pallidum?
2. Another interesting remark regarding the granule stage of
spirochaetes may be found in Sir Patrick Manson’s well-known book
on Tropical Diseases. In the last (fifth) edition, published in 1914,
on page 225, regarding the etiology of relapsing fever, he writes:
‘Obermeier and von Jaksch describe certain refractile bodies present
in the blood during the fever intermissions. The latter author says
that he has observed the development of these bodies into short rods,
from which the typical spirochaetes are eventually evolved.’ These
remarks have also appeared in earlier editions of the book.
3. Dutton and Todd, writing in 1905 concerning some of their
experiments on the nature of human tick fever in the Congo Free
State, record that: ‘ In some preparations of stomach or malpighian
tubules [of spirochaete-infected ticks] no parasites were at first seen;
but if a little human serum, taken from one who had never had tick
fever, were added, in from 8 to 24 hours the preparations became
fairly crowded with spirochaetes.* These observations may be
explained by spirochaetes passing through a granule phase. .
402
VIII. THE SPIROCHAETES OF THE HUMAN MOUTH
Two species of spirochaetes were recorded as occurring in the
human mouth about forty years ago. These are S. buccalis,
Steinberg (often ascribed to Cohn, 1875), and 5 . dentium , Miller
(often attributed to Koch, 1877). The former is said to be longer
and thicker than the latter. The smaller spirochaete is also the
more flexible. According to Hartmann and Miihlens (1906),
S. dentium in culture measures 4 /* to 12 /* in length by 0*3/1 to
0*6 fi in breadth, or less if stains other than Loffler's are used, while
S. buccalis is 12 p to 20 /* in length by 0*5 fi to 1 /* in breadth.
Hoffmann and Prowazek (1906) published microphotographs of
specimens of 5 . dentium about 5 /* to 7*5 /* long, and of
5 . buccalis about 8*5 n to 14*5 ft long. Intermediate types also
occur, as was shown by these authors, such forms being named
Treponema intermedium by Dobell (1912). Hartmann and
Miihlens did not see much internal structure in S. dentium , but in
5. buccalis they record the presence of an ‘ undulating * membrane in
some specimens. Miihlens (1907) figured stained specimens of
5 . buccalis and S. dentium in which chromatin coloured granules
were distributed along the bodies of the organisms. His figures of
5. dentium, as measured by me, are about 4 fi to 7/1 long and of
5 . buccalis about 15 fi to 23 /*.
Noguchi (1912) has succeeded in cultivating a number of species
of spirochaetes from the human mouth. He places these in the genus
Treponema , making three new species, namely, T. macrodentium ,
T. microdentium and T . mucosum , but they cannot be easily
distinguished morphologically. The last-named was obtained from
cases of pyorrhea alveolaris. They all differ from S. buccalis.
In the course of my researches I have observed the parasites
ascribed to Cohn and to Koch, these being the two common spiro¬
chaetes seen in the mouths of natives of the Sudan and of Europeans
in England, as well as the forms described and cultivated by later
investigators. Some of the mouth spirochaetes are not very active,
but there is marked corkscrew and boring movement, and they are
flexible. Tangles or tomenta of these mouih spirochaetes are common.
Internal structure is seen with some difficulty, but in some specimens
it can be determined, and chromatin granules are then observed.
4°3
S. dentium (h gs. 53-57) has tapering ends, and varies in length from
4 fi to 10 fi, as found by measuring 40 specimens from some of
my own preparations. S. dentium is so small that few details of its
structure can be determined, and in consequence, it might be placed
by some authorities in the genus Treponema. S. buccaiis
(figs. 58-68) has somewhat rounded or bluntly acuminate ends, and
varies in length from 9 fi to 22 /1, as determined from 110 specimens
measured. A slight membrane or crest may sometimes be observed
in the latter species (figs. 60, 65). Intermediate forms were found,
which might be considered to connect the two species.
The spirochaetes of the mouth take up stains well and with
relative ease. Intracellular stages, if they occur at all, are
uncommon. Multiplication by binary fission has also been observed.
Coccoid bodies or granule stages of the mouth spirochaetes are
formed, but appear to be relatively few in number.
No marked morphological difference has been observed between
the spirochaetes occurring in the mouths of Europeans in England
(four cases examined), and in natives of the Sudan (eight cases
examined). However, it may be of interest to note that in the Sudan
the predominant spirochaete in the mouths of the natives whom
I examined was S. buccaiis , S. dentium being relatively uncommon.
Spirochaetes that were either 5 . dentium or closely allied thereto
were found in the mucus from the pharyngeal and laryngeal regions
of a normal monkey in the Sudan.
A spirochaete of the human throat, often associated with
fusiform bacilli, may be mentioned here. Spirochaeta vincenti
occurs in the throat in certain conditions, such as Vincent’s angina.
It has recently been described by Drs. J. G. and D. Thomson (1914).
It is said to be an elongate spirochaete, with a flexible, irregularly
coiled body, tapering at both ends. The spirochaete, according to
the present state of our knowledge, does not seem to be identical with
5 . bronchialiSy differing from it morphologically and in the situations
in which it is found. S. vincenti was named by Blanchard in 1906, but
as no description of the spirochaete was given at the time of naming,
the species name is probably not valid. Little is known of the
range of morphological variation of S. vincenti. I have measured
those drawn by Thomson and find that they are from 10 fi to 18/1
long, while the few figured by Miihlens (1907) I find to measure
9 Hj io (i and 23/1. Casteilani and Chalmers state that it is 12 ^
to 25 n long. It is possible that this species will have to be merged
into another one, such as 5 . bronchialis, when it has been more fully
investigated.
IX. THE SPIROCHAETES OF THE HUMAN MOUTH CONTRASTED
WITH S. BRONCHIAUS
There are general differences between these two groups of
spirochaetes, and these differences may be set forth with a view to
aiding those workers in the tropics who may meet with cases of
bronchial spirochaetosis. However, the said differences must not be
emphasised too much or asserted dogmatically, as further work may
lead to modifications.
The bronchial spirochaetes appear to be more active than those
of the mouth.
5 . bronchialis dies very rapidly outside the respiratory tract.
This has also been noticed by Chalmers and OTarrell and by
Taylor. Oral spirochaetes, on the other hand, can live for some
hours outside the human mouth.
Bronchial spirochaetes, as a rule, stain with difficulty and not so
easily as those obtained from the mouth. The former are much more
fragile and slender than 5. buccalis.
Coccoid bodies are frequently produced by 5 . bronchialis, and
are probably the cross-infective stage of the parasite. Coccoid
bodies appear to be less frequently formed by oral spirochaetes.
Intracellular stages of S. bronchialis are occasionally seen. No
such stages have been observed in the cases of spirochaetes from the
mouth, which I have examined.
Tangles are not usually formed by 5 . bronchialis, but are
commonly produced by numbers of spirochaetes aggregating in
clusters in the oral cavity.
X. MODE OF INFECTION
The situation in which Spirochaeta bronchialis is found largely
precludes its dissemination direct from man to man by the aid of
insects, though they may serve as indirect transmitters of the
parasite.
It is highly probable that man himself acts as the reservoir of the
virus, and that direct contact of infected with healthy persons is
responsible for the spread of bronchial spirochaetosis. The spray
exhaled with the expired air appears to be contaminated with the
resistant coccoid bodies produced by S. bronchialis. Such fine spray
inhaled by an uninfected person whose bodily resistance is somewhat
lowered—as after a chill—results in an attack of bronchial
spirochaetosis. It is possible that spirochaetes as such might be
exhaled in the spray, but as has been pointed out they are fragile
and quickly die outside the body. It is also possible that the
inhalation of dust contaminated with dry sputum from patients
suffering from bronchial spirochaetosis may excite an attack, while
the nasal secretions passed on to linen and allowed to dry, the linen
then being packed indiscriminately with other soiled clothing, may
serve to transmit the parasite to new hosts.
The agency of insects in spreading bronchial spirochaetosis is
somewhat remote, but it must be remembered that certain flies are
partial to sputum, which they not only ingest but which may soil
their bodies and feet. While the frail, motile spirochaetes
are not likely to be carried as such in or on the bodies of flies, it is
quite possible that the more resistant coccoid bodies may be carried
about by flies, and deposited by them either directly on the lips or
nose of man, or in foods such as milk, whence they may reach the
pharynx and perhaps pass downwards into the trachea and bronchi.
However, the direct contamination of healthy persons by infected
ones is at present much the most likely means of dissemination of
5 . bronchialis .
XI. GENERAL REMARKS
Certain points of interest connected with Spirochaeta bronchialis
and bronchial spirochaetosis may now be considered.
That Spirochaeta bronchialis is disease producing is undoubted.
The organisms have been found in cases of chest complaints,
especially those with bronchitic symptoms, where no other cause for
the affection was assignable. S. bronchialis was frequently the only
organism present in the cases in the Sudan examined by me.
Examination of a series of preparations from several chronic cases,
406
each considered separately, has shown that, on most of the occasions
on which sputum was examined, bacteria of any kind were either few
or absent. Sometimes organisms identical morphologically with
Diplococcus ( Micrococcus ) catarrhalis were found on a single
preparation of a series extending over a number of days, but this was
uncommon. Occasionally bacteria were found in smears of mouth
spirochaetes. On still rarer occasions cells from the lungs were
observed in which endocellular spirochaetes and coccoid bodies
related thereto were present in company with diplococci (fig. 52).
The contrast between the coccoid bodies and the bacteria, both in
size, morphology and staining reactions, was always noticeable. In
one case only were pneumococci found associated with Spirochaeta
bronchialis , the patient being a European soldier who might have
become infected with spirochaetes after a chill had favoured
pneumonic symptoms. No mycological agent has been found in any
of my preparations. Chalmers and O’Farrell also have had a similar
experience, and they state that the fungi responsible for broncho-
moniliasis, broncho-nocardiasis, broncho-aspergillosis, broncho-
penicilliosis, broncho-mucor-mycosis, broncho-sporotrichosis, have
not been found by them 4 in the sputum of their cases of bronchial
spirochaetosis either by direct examination or by cultivation.*
Further, Chalmers and O’Farrell succeeded in experimentally
inducing bronchial spirochaetosis in a monkey, the parasites and
symptoms closely resembling those in man (cf. figs. 19, 49).
It is also of interest to note that Chalmers and O’Farrell, and
Taylor, both writing in 1913, arrived at the same conclusions
regarding bronchial spirochaetosis in Khartoum and Uganda
respectively. In each district S. bronchialis is held to be the cause
of the haemorrhagic bronchitis noticed. Also each of the above
workers state that there is an abundance of S. bronchialis during the
attack, that the decrease in numbers synchronises with the abatement
of symptoms, and that the spirochaetes disappear from the sputum
entirely or are found with great difficulty after a few days. All
these facts have a distinct significance when considering the etiology
of the disease.
5 . bronchialis I find to be morphologically different from the
spirochaetes of the oral cavity, as mentioned previously. It is an
entity in itself, and the malady caused by it also appears to be
distinctive. Its pathogenic action also seems to be unlike that of the
debatable S. vincenti of Vincent’s angina, in which a throat
membrane may at times be produced.
S. bronchialis has a somewhat extensive though scattered
distribution. Bronchial spirochaetosis has been reported from
Europe, the cases being discussed by Chalmers and O’Farrell. It
has also been recorded from Ceylon, India, the Philippines, and the
West Indies. In East Africa the disease has been reported from the
Anglo-Egyptian Sudan and Uganda. In West Africa the disease
has been recently notified from various parts of the Gold Coast
Colony. Spirochaeta bronchialis has thus been reported from four
continents and from different districts in each. This scattered but
wide distribution of the parasite suggests that it may be a very
general but easily overlooked organism, of far more frequent
occurrence than is usually supposed.
For the benefit of medical men who are interested in the subject
and to whom Chalmers and O’Farrell’s paper is not available, the
treatment recommended by them is here quoted : —
1 The first essential is rest in bed, good food, and ventilation.
The second is arsenic in some form, preferably associated with
glycerophosphates. These may be given by the mouth with excellent
results, or intramuscularly as an injection of: —
Sodium cinnamate . 0 05 grm.
Sodium cacodylate . o’10 ,,
Sodium glycerophosphate . 0*10 ,, ’
Taylor, in Uganda, prescribes ‘ Arsenious acid by the mouth in
increasing doses.’
At the time when these treatments were recommended (end of
1913) they had been employed in relatively few cases, but had given
satisfactory results.
Chalmers and O’Farrell, and Taylor also discuss the
co-existence of bronchial spirochaetosis with other chest diseases,
such as lobar pneumonia, details of which should be sought in the
original papers. Bronchial spirochaetosis may be suspected in
atypical cases of pneumonia and bronchitis, and may be mistaken
for incipient phthisis.
XII. SUMMARY AND CONCLUSIONS
1. Spirochaeta bronchialis is an organism presenting marked
polymorphism, a feature that has only been determined by the
examination of numerous preparations from various patients. It
produces bronchial affections in the Sudan and in other parts of the
world.
2. 5 . bronchialis , as investigated in the Anglo-Egyptian Sudan,
varies in length from 5 fi to 27 fi 9 and its breadth is about 0*2 p to
0 6 fi . These variations are due to the processes of growth and
division. Many of the parasites measure either 14 fi to 16 fi long,
or 7 fi to 9 fi , the latter resulting from transverse division of the
former. The ends show much variation in form, but approach the
acuminate type on the whole. The discrepancies in dimensions
given by the very few previous workers on S. bronchialis are the
result of the measurement of a limited number of parasites. All such
sizes can be found on some occasion during the progress of the
disease, when a larger number of spirochaetes is examined.
3. The movements of S. bronchialis are active, but of relatively
short duration, when it is removed from the body. The number of
coils of the spirochaetes is rather an index of its rapidity of motion
than a fixed characteristic of the species.
4. The motile phase of S. bronchialis is succeeded by one of
granule formation, the granules or coccoid bodies serving as a resting
stage from which new spirochaetes are produced. The formation of
coccoid bodies and the reproduction of spirochaetes from them can
be observed in life.
5. S. bronchialis is a species distinct from the spirochaetes
occurring in the mouth. It differs from them in morphology,
pathogenicity and in staining reactions. It is not a developmental
form of any bacterium, and is an entity in itself.
6. The passage from man to man is effected most probably by
means of spirochaetes, and especially coccoid bodies, that leave the
body in the spray with expired air and by way of the nasal
secretions. Owing to the fragility and short life of S. bronchialis
extracorporeally, the resistant coccoid bodies in air, dried sputum
and dust, and possibly also on the bodies of flies and other insects,
are probably instrumental in inducing attacks of bronchial spiro-
4°9
chaetosis in human beings, especially those having a lowered bodily
resistance, such as after a chill.
7. Spirochaetes of the type of S. dentium , measuring 4 fi to 10 fi
in length, have been found in mouths of natives of the Sudan and in
those of English people in England. Also S. buccalls , measuring
9 fi to 22 fiy has been obtained from the same sources.
8. S. bronchialis will probably prove to be of more frequent
occurrence than is known at present.
REFERENCES
Relating to Bronchial Spirocbaetosis
Castellani, A., and Chalmers, A. ]. (1913). Manual of Tropical Medicine, 2nd edition,
pp. 402 and 1283.
Chalmers, A. J., and O’Farrell, W. R. (1913). Bronchial Spirochaetosis. Journ. Trop.
Med. and Hyg ., Vol. XVI, pp. 329-334.
Chamberlain, W. P. (1911)* The Occurrence in the Philippines of Associated Spirochaetae
and Fusiform Bacilli in Ulcers of the Throat (Vincent’s Angina), of the Mouth, and
of the Skin, and in Lesions of the Lungs (Bronchial Spirochaetosis). Philippine Journ.
Science , B, Vol. VI, pp. 489-498. 2 Pis.
- (1914)* Spirochaetae and Fusiform Bacilli in various lesions in the Philippines. Amer.
Journ. Trop. Dm., Vol. II, pp. 246-255. 1 PI.
Harper, F. S. (1914). Bronchial Spirochaetosis. Journ. Trop. Med. and Hyg Vol. XVII,
p. 194.
Macfie, J. W. S. (1915). Bronchial Spirochaetosis. Journ. Trap. Med. and Hyg., Vol.
XVIII, pp. 63-65.
Taylor, J. A. (1914). Bronchial Spirochaetosis in Uganda, with pneumonic symptoms.
Annals Trop. Med. and Parasitcl ., Vol. VIII, pp. 13-18.
- (1914)* Bronchial Spirochaetosis in Uganda. Annual Med. and Sanit. Rept., Uganda
Protectorate , for 1913, pp. 80-86.
Relating to Spirochaetes of the Human Mouth
Hartmann, M., and MOhlens, P. (1906). Untersuchungen iiber Bau und Entwicklung der
Zahnspirochaten. Zeitscbr. f. Hyg. und Infektionskrankb., Bd. LV, pp. 92-112. 2 Pis.
Hoffmann, E., and Prowazek, S. von (1906). Untersuchungen iiber die Balanitis- und
Mundspirochaten. Centralbl. /. Bakt ., 1 Abt., Orig., Bd. XLI, pp. 741-744, 817-821.
1 PI.
MOhlens, P. (1907). Vergleichende Spirochatenstudien. Zeitscbr. f. Hyg. und Infektions¬
krankb., Bd. LVII, pp. 405-416. 3 Pis.
410
Noguchi, H. (1912). Cultural Studies on Mouth Spirochaetae £ Treponema microdentivm
and macrodentium). Journ. Exper. Med., Vol. XV, pp. 81-89. 5 Pis.
- (1912). Treponema mucosum (new species), a mucin-producing spirochaeta from
pyorrhea alveolaris, grown in pure culture. Journ. Exper. Med., Vol. XVI, pp. 194-198.
1 PI.
Thomson, J. G., and Thomson, D. (1914). Some Researches on Spirochaetes occurring in
the Alimentary Tract of Man and some of the Lower Animals. [Marcus Beck Lab.
Repts.—No. 3.] Proe. Roy. Soc. Med., Vol. VII, Pt. 1, pp. 47-70. 2 Pis.
Relating to Spirochaetes in General
Blanchard, R. (1906). Spirilles, Spirochetes et autres microorganismes A corps spirals.
Arch, de Parasitologic, Vol. X, pp. 129-149.
Bosanquct, W. C. (1911). Spirochaetes. Pp. 152. W. Saunders Company, Philadelphia
and London.
Dobell, C. C. (1912). Researches on the Spirochacts and related Organisms. Arch. /.
Protistenkunde , Bd. XXVI, pp. 117-240. 5 Pis.
Dutton, J. E., and Todd, J. L. (1905). The Nature of Human Tick-Fever in the Eastern
Part of the Congo Free State. Liverpool Scb. Trap. Med., Memoir XVII. See p. 17.
Fa nth am, H. B. (1907). Spirochaeta {Trypanosoma) balbianii (Certes), its Movements,
Structure, and Affinities j and on the Occurrence of Spirochaeta anodontae (Keysselitz)
in the British Mussel, Anodonta cygnea. (Preliminary Account.) Ann. & Mag. Nat.
Hist., ser. 7, Vol. XIX, pp. 493-501.
- (1907). The Movements of Spirochaetes, as seen in S. balbianii and S. anodontae.
Brit . Assoc. Rept., 1907 (Leicester), pp. 554-355.
- (1908). Spirochaeta (Trypanosoma) balbianii (Certes) and Spirochaeta anodontae
(Keysselitz) : their Movements, Structure, and Affinities. Quart. Journ. Microsc. Sci.,
Vol. LII, pp. 1-73. 3 Pis.
-(1908). The Spirochaetes : A review of some border-line organisms between animals
and plants. Science Progress, Vol. Ill, No. 9, pp. 148-162. 4 Text-figs,
- (1909). The Spirochaetes found in the Crystalline Style of Tapes aureus: a Study
in Morphological Variation. Parasitology, Vol. II, pp. 392-408. 1 PI.
-(1911). Some Researches on the Life-Cycle of Spirochaetes. Annals Trop. Med.
and Parasitol., Vol. V, pp. 479-496. 6 Text-figs.
- (1914). The Granule Phase of Spirochaetes. Annals Trop. Med. and Parasitol.,
Vol. VIII, pp. 471-484. 2 Text-figs.
Fantham, H. B., and Porter, A. (1909). The Modes of Division of Spirochaeta recurrentis
and 5 . duttoni as observed in the Living Organisms. Proc. Roy. Soc., B, Vol. LXXXI,
pp. 500-505*
Gross, J. (1910). Cristispira nov. gen. Ein Beitrag zur Spirochatenfrage. Mittb. a. d. zool.
Station zu Neapel , Bd. XX, pp. 41-93. 1 PI.
- (1912). Ueber Systematik, Struktur und Fortpflanzung der Spironemacea. Centralbl.
/. Baku , I Abt., Orig., Vol. LXV, pp. 83-98.
Noguchi, H. (1913). On some of the Recent Advances in the Field of Microbiology; with
Demonstrations of the Pure Cultures of various Spirochaetes, of the Viruses of Rabies
and Poliomyelitis, and of Treponema pallidum in the Brains of General Paralytics.
[Occasional Lectures.] Proc. Roy. Soc. Med. y Vol. VII, Pt. 1, pp. 3-30. (See pp. 5, 19.)
Zuelzer, M. (1911). Ueber Spirocbaeta plicatilis Ehrbg. und dcren Verwandtschafts-
beziehungen. Arch. f. Protistenkunde , Bd. XXIV, pp. 1-59. 4 Pis.
412
EXPLANATION OF PLATE XXXIV
All figures were outlined with an Abb6-Zeiss camera lucida, using a
Zeiss 2 mm. apochromatic objective and compensating ocular 18. The
magnification is approximately 2,600 diameters.
Figs. 1-39. Spirochaeta bronchialis from the Anglo-Egyptian Sudan. Trophic
stage.
Figs. 1-6. Small, young forms of S. bronchialis.
Figs. 1,3. Spirochaetes with both ends slightly differing in form.
Figs. 2, 4. Forms with both ends alike. In fig. 2 the edge of the membrane
shows as a line on the body.
Figs. 6-13. S. bronchialis . Growth forms showing variation in thickness and
length.
Fig. 10. Spirochaete with relaxed membrane.
Fig. 13. Spirochaete drawn from preparation obtained from Kodok, Sudan.
Figs. 14-24. Longer forms.
Fig. 16. Form with slight membrane.
Figs. 19, 23. Forms with many small coils, fixed when in rapid motion.
Fig. 19 drawn from specimen from an experimentally infected
monkey.
Figs. 14, 20. S. bronchialis from a case at Kodok.
Fig. 22. The edge of the membrane (crista) shows as a wavy line on the
body.
Figs. 25-35. Some of the commoner (‘average’) types of S. bronchialis ,
showing variation in the number of coils.
Figs. 36-39. Some of the largest forms of 5 . bronchialis encountered.
Fig. 36. Spirochaete with slight membrane.
Fig. 37. Form with one end more pointed than the other.
Fig. 38. Very broad form.
Fig. 39. Long form with slight membrane.
Figs. 40-45. Spirochaeta bronchialis from Uganda.
Figs. 40-44. Trophic forms showing the same appearances and structure
as the average types of S. bronchialis occurring in the Sudan.
Fig. 45. Parasite showing the formation of coccoid bodies.
Figs. 46-52. The granule phase of S. bronchialis from cases in the Sudan.
Figs. 46-49. Free spirochaetes containing coccoid bodies. Fig. 49 drawn
from a preparation obtained from an experimentally infected
monkey.
Fig. 50. Coccoid bodies in process of liberation at one pole of the spirochaete.
Fig. 51. Cluster of small coccoid bodies with young spirochaetes emerging
from the group.
Fig. 52. Part of cell from the air passages, containing two spirochaetes, a
group of coccoid bodies retaining the arrangement of the spiro¬
chaete from which they were liberated, and two diplococci.
Figs. 53-57. Spirochaeta dentium from mouths of Europeans and natives of the
Sudan.
Figs. 53, 54. S. dentium , from natives of the Sudan.
Figs. 55, 56. 5 . dentium , from Case 1, Cambridge.
Fig. 57. S. dentium , from Case 2, Cambridge.
Figs. 58-68. Spirochaeta buccalis from mouths of Europeans and natives of
the Sudan.
Figs. 58, 62, 63, 65. S. buccalis , from Case I, Cambridge. Fig. 65 shows a
membrane or crista.
Fig. 59- S. buccalis , from Case 2, Cambridge.
Fig. 64. S . buccalis , from Case 1, Liverpool.
Figs. 60, 61, 66-68. S. buccalis , from natives of the Sudan. In fig. 60 a
slight membrane is indicated.
Annals Trap. MeJ. & Parasite!., Pol. IX
PLATE XXXIP
SPIROCHAETA BRONCHIALIS, Figs. 1—52
SPIROCHAETES FROM HUMAN MOUTH, Figs. 53—68
4'3
NOTES ON CERTAIN ANIMAL
PARASITES OF DOMESTIC STOCK IN
SIERRA LEONE
•
\Being the Fourth Report of the Thirty-second. Expedition of the
Liverpool School of Tropical Medicine, 1914-15.]
BY
WARRINGTON YORKE
AND
B. BLACKLOCK
(Received for publication 29 June, 1915)
SERIOUS EPIDEMICS OF DISEASE AMONGST CATTLE IN
SIERRA LEONE
From time to time heavy losses have been experienced by cattle
owners in Sierra Leone owing to epidemics, during which large
numbers of cattle die.
In December, 1914, we were requested by the Administration to
investigate the nature of a serious epidemic which had broken out
amongst cattle in the northern part of the country. The cattle in
question had been brought down originally from French Senegal;
they were practically all bullocks, owing to restrictions respecting
the export of cows put into force by the French Administration.
We proceeded to Batkanu, the capital of the Karene district, but
were informed on our arrival that the epidemic had ceased and fhat
the animals were no longer dying. We found 90 cattle in the
Government Warri, all of which appeared to be in a healthy
condition. Most of the animals were in the Karene district during
the epidemic, and had remained apparently uninfected; a few,
however, had been ill, and seemed to have recovered. Whether
these animals had really suffered from the infection which had
resulted in the death of large numbers of others is uncertain, but
Dr. Clearkin, the medical officer in charge of the district, was of the
opinion that this was the case. We were informed that the epidemics
occurred towards the end of the rainy season, and that they
coincided with the appearance of large numbers of a fly {Stomoxys
nigra), which was held responsible. This hypothesis is in
accordance with the history given by the natives; those questioned
by us stated that the disease appeared every five or six years,
during the rainy season, and that it was due to the bites of the fly
(Stomoxys). It was believed that the infection was contracted by
the animals whilst being brought down from French country, where
cattle breeding is carried on extensively.
The course of the disease was rapid, the symptoms being fever,
staring coat, emaciation, and death in a month or six weeks.
Fresh and stained preparations of blood of each of the 90 cattle
in the Warn were examined, and rats were inoculated with blood
from every tenth animal. As a result of this examination four
cattle were found to be infected with trypanosomes; two with
T r vivax r, one with T . congolense and one with T. gambiense. In
two of the animals Piroplasma bigeminum was found, and in about
30 per cent, a parasite belonging to the genus Theileria. Two
healthy oxen were inoculated with blood from each of the animals
suffering from infection with T . vivax . The animals inoculated
from the first ox remained healthy, and trypanosomes were never
seen in their blood; those inoculated from the second, which also
harboured Piroplasma bigeminum in its blood, became infected both
with T. vivax and with Piroplasma bigeminum , but exhibited no
signs of disease, and were alive and in good condition when we left
the colony three months later.
In view of the fact that we did not see these animals until the
epidemic had ceased, it was impossible for us to make any post¬
mortem examination, and there was no material at our disposal
upon which a definite conclusion could be reached as to the cause of
the great mortality which had occurred. Although the examination
of blood proved that the cattle were infected with several protozoa,
any one of which is potentially the cause of epidemics fatal to cattle,
it would not be possible to make any definite statement on the
subject until observations can be made on the spot during the time
when the epidemic is running its course, so that animals can be
examined systematically, the clinical course of the disease watched
and material obtained immediately after death. We cannot
emphasise too much the importance of properly directed and
sustained work in the investigation of problems of such economic
importance; it is only by this means that the true nature of these
ever-recurring epidemics will be elucidated.
TRYPANOSOMIASIS
During our visit to Sierra Leone we had the opportunity of
examining a considerable number of domestic stock, both in the
Colony and in the Protectorate. In all, 143 cattle were examined,
and trypanosomes were found in nineteen. The proportion of
infected cattle must, without doubt, be very much greater than this,
as in the majority of cases an examination of a single fresh
preparation of the blood was all that could be undertaken. These
cattle, which were brought to Freetown for slaughter, appeared to
be the most heavily infected; trypanosomes were discovered in the
blood of fourteen out of thirty-four animals immediately after they
had been killed at the Military slaughter-house. A probable
Explanation of this is that so soon as animals begin to lose condition
they are taken by the natives to be sold in the Freetown market.
In the nineteen infected animals Z. congolense was found eleven
times, Z. vivax twice, a double infection of T. congolense and
Z. vivax five times, and T. gambiense once.
The blood of ten dogs was examined, but in no cases were
trypanosomes found. One of the animals was, however, proved to
be infected with Z. congolense by inoculation of rats. No instance
of infection with trypanosomes was found in the seven goats and
seven sheep examined.
In connection with the information derived from examination of
domestic stock it is of interest to refer to observations on the amount
of trypanosome infection obtaining in G. pal falls.
Four hundred wild Glossina palpalts caught on the Cape
Lighthouse Peninsula were dissected and examined; trypanosomes
were found in twenty-one of them.
4i 6
Tabli I.—Showing the manner in which trypanosomes were recognised in infected domestic
stock
No.
Animal
Trypanosomes
found in
peripheral
blood
Animals
inoculated
Result of
inoculation
Diagnosis
i
Ox 11
T. congolense
...
...
7 . congolense
z
Ox 23
T. vivax
Ox I
Ox II
Rat 15a
Rat 15b
Rat 15c
Negative
Negative
Negative
Negative
Negative
T. vivax
3
Ox 88
7 . congolense
Rat 21
7 . congolense
T. congolense
4
Ox 89
T. vivax
Ox III
Ox IV
Rat 16a
Rat 16b
Rat 16c
7 . vivax
7 . vivax
Negative
Negative
Negative
7 . vivax
5
Ox 100
Negative
Rat 24
T. gambiense
T. gambiense
6
Ox 108
T. congolense
...
...
T. congolense
7
Ox in
T. congolense
7 . congolense
8
Ox 112 ...
T. congolense
T. congolense
9
Ox 120
T. congolense
T. congolense
IO
Ox 121 ...
T. congolense
...
T. congolense
n
Ox 137 ...
(T. congolense
|T. vivax
Rat 11a
Rat nb
Negative
Negative
J T. congolense
\ T. Ct« 7 A*
12
Ox 140
7 . congolense
Rat 12a
Rat 12b
7 . congolense
T. congolense
T. congolense
13
Ox 141 ...
7 . congolense
Rat 13a
Negative
7 . congolense
14
Ox 142 ...
7 . congolense
Rat 13b
Negative
7 . congolense
15
Ox 143 ...
7 . congolense
...
...
7 . congolense
16
Ox 144 ...
( 7 . congolense
(T. vivax
j 7 . congolense
( 7 . vivax
17
Ox 145 ...
| T. congolense
( 7 . vivax
Mi
...
( 7 . congolense
( 7 . two*
18
Ox 146
f T. congolense
\T.vivax
f 7 . congolense
\ 7 . vivax
19
Ox 147 ...
( T. congolense
{ 7 . vivax
...
f 7 . congolense
( 7 . vivax
20
Dog 155 ...
Negative
Rat 30a
Rat 30b
7 . congolense
T. congolense
7 . congolense
4 ! 7
Table 2. —Giving results of dissection of wild Glossina palpalis found to be infected with
trypanosomes
No. of fly
Salivary
Proboscis
Intestine
Result of inoculation
glands
«
0
0
4 - 4 - 4 -
2 rats, negative
2
0
+ 4 -
0
3
0
4 - 4 -
4 - 4 -
1 rat, gut and proboscis contents,
negative
4
0
4 - 4 - 4 -
0
5
0
4 - 4 -
0
6
0
4 - 4 -
0
7
0
4 - 4 - 4 -
0
8
0
4 -
0
(1 rat, gut contents, negative
9
0
4 - 4 - 4 -
4 - 4 - 4 -
J 1 rat, proboscis contents, died on 6th
l
10
0
4 -
0
11
0
4 - 4 - 4 -
0
12
0
4 - 4 - 4 -
0
13
0
4 * !
0
14
0
4 - 4 - 1
0
>5
0
0
4 -
(i rat, gut contents, died on 6th day
16
0
4 - 4 - 4 -
4 - 4 - 4 -
j 1 rat, proboscis contents, died on 6th
[ day
>7
0
4 -
0
18
0
4 -
0
>9
0
4 -
0
20
0
4 - +
4 - 4 - 4 -
(i rat, gut contents, negative
{1 rat, proboscis contents, negative
21
0
4 -
0
4 * Signifies trypanosomes scanty.
4- 4 - Signifies trypanosomes numerous.
4 - 4 * 4 * Signifies trypanosomes swarming.
It will be seen from Table 2 that of the twenty-one flies found
to contain trypanosomes, the proboscis alone was involved in
fifteen, the gut alone in two, whilst there was a heavy infection of
both gut and proboscis in four. No instance of invasion of the
salivary glands was encountered. Assuming that these parasites
were trypanosomes pathogenic to man and domestic stock, the result
of previous work allows us to conclude that fifteen of the above flies
were infected with T . vivax , four with T . congolense , and the
remaining two with a trypanosome belonging either to the
Congolense or the Gambiense group; which, we are unable to
decide owing to the fact that the cycle of development of the
trypanosome in the fly was incomplete. Inoculations into rats were
made, with negative results in each case, from five of the flies
found to be infected. Unfortunately in two instances (Flies 9
and 16) the rats which had received the contents of the proboscis
died six days after inpculation, but in two other cases (Flies 3
and 20) the rats lived long enough to have become infected. The
fact that these animals did not become infected does not prove that
the trypanosome found in the flies was not T. congolense , as it has
now been established that it is always easy to infect rats with this
trypanosome. A similar failure of T. congolense to establish itself
in these animals is seen in the case of three rats inoculated
respectively from Oxen 137, 141 and 142 (see Table I), which were
found on direct examination of the blood to be suffering from an
infection with T. congolense . Previously the trypanosome in
question would have been designated T . nanum y but recent
work shows that the difference between the latter parasite and
T. congolense is only an apparent one.
A small number (95) of wild G. palpalis was fed on rats; no
infection resulted.
BABESIASIS AND THEILERIASIS
Whilst examining blood films of cattle, species belonging to the
genera Babesia and Theileria were seen. In about 5 per cent, of
the animals examined a few large double pear-shaped bodies and
an occasional large amoeboid form were encountered in the red cells.
This parasite was undoubtedly Piroplasma bigemtnum.
+i 9
The other parasite occurred much more frequently, being found
in between 20 and 30 per cent, of the animals examined. Small rod-
and ring-shaped bodies showing distinct cytoplasm and chromatin
were seen in the red cells. In some animals 1 or 2 per cent, of
the cells were invaded, whilst in others only an occasional parasite
was found. These bodies seem to be identical with those discovered
by Macfie (1914) in cattle in Nigeria. They did not appear to
produce any symptoms of disease. Most of the animals in which
they were found were in perfect health; a few were emaciated, but
in these trypanosomes were also found.
Owing to the fact that so large a proportion of cattle were
discovered to harbour these bodies in their blood—usually they
were present in very small numbers only—we did not consider that
inoculation experiments into local cattle would afford any evidence
as to the inoculability or otherwise of the parasite. No 1 blue bodies *
of Koch were found in the blood, or in smears made from the
kidneys, liver, spleen, lungs, or bone marrow of six infected oxen.
Theiler (1907) describes Theileria parva and Theileria mutans as
being very similar in appearance. He differentiates between them
on the following grounds. Th. parva is very virulent, is non-
inoculable, and is characterised by the presence of the ‘ blue bodies 1
of Koch; whereas Th. mutans is non-virulent, is inoculable, and the
‘blue bodies' of Koch are not found. There is little reason to
doubt, therefore, that the parasite in question is Theileria mutans.
All the ticks which could be found were collected from each of
the ninety cattle examined in the Warri at Batkanu; the animals
were thrown and carefully searched. The only species encountered
were Boophilus australis (126 adults, and 395 nymphs) and
Amblyomma variegatum (4 adults, and 28 nymphs). We are
indebted to Professor G. H. F. Nuttall for kindly determining
them.
CONCLUSIONS
I. Cattle are not bred to any extent in the Colony of Sierra
Leone or in most parts of the Protectorate. Bullocks are, however,
imported from French Senegal in considerable numbers, and
gradually find their way down to Freetown for slaughter.
420
2. Serious outbreaks of disease in the form of epidemics occur
amongst the cattle of Sierra Leone.
3. These epidemics have been attributed to various causes, but
their real nature is still obscure.
4. Trypanosomiasis of cattle is common. Of the thirty-four
animals examined at the slaughter-house in Freetown, fourteen
(41 per cent.) were found to be infected. As only a single blood film
was examined, the real percentage of infections is certainly much
higher.
5. T. congolense and T. vivax are the parasites most commonly
found. T. gambiense was met with once.
6. About 5 per cent, of the animals examined were found to be
infected with Piroplasma bigeminutn.
7. Theileria mutans was encountered in between 20 and 30 per
cent, of the cattle examined.
REFERENCES
Macfie, J. W. Scott (1914). Notes on some Blood Parasites collected in Nigeria. Annals
Trop . Med . and Parasitol ., VIII, pp. 439-468.
Theilek, A. (1905-6). Report of the Government Veterinary Bacteriologist. Transvaal
Department of Agriculture, Pretoria.
- (1906-7). Report of the Government Veterinary Bacteriologist. Transvaal Depart¬
ment of Agriculture, Pretoria.
421
THE ETIOLOGY OF JUXTA-ARTICULAR
SUBCUTANEOUS NODULES
BY
J. B. DAVEY, M.B. Lond., D.T.M. Liverpool
(Received for publication 16 June, 1915)
Whilst engaged, during the months of June, July and August,
1913, in examining the natives of the low-lying part of the Dedza
district of Nyasaland for sleeping sickness, two facts soon forced
themselves upon my attention. One of these was the prevalence of
yaws and the other the frequent occurrence amongst these people
of firm subcutaneous nodules, varying in size from that of a pea to
that of a duck’s egg, in the vicinity of joints. Observations were,
therefore, made on a series of 2,378 adults (964 males and 1,414
females) and 567 children, natives of this district, and the results are
presented in this paper, as they seem to be of interest in elucidating
the etiology of such nodules.
The nodules which, as I have said, vary considerably in size,
are firm, painless, not tender, show no tendency to break down,
and are always situated in close relationship to the subcutaneous
portions of bone. They are, however, freely movable on the bones
and the skin over them, except in a few cases where definite signs
of trauma accounted for adhesion to the skin. Their favourite site
is the posterior border of the ulna, about two inches from the tip of
the olecranon, and they were found, in this situation, on one or
both sides, in 55 of the 2,378 adults examined. Other fairly
common sites are the great trochanter of the femur and the lower
part of the patella: less commonly they were found over the
malleoli, and in one case on the zygoma. Frequently several were
found in one individual at these various positions. At the patellar
site they were usually accompanied by enlarged bursa. Altogether
in 80, out of the 2,378 adults examined, they were found in one or
more positions. They were never found in children, of whom 567
were examined for them.
4 22
These observations led me to the conclusion that the nodules are
those referred to, in works on Tropical Medicine, as ‘ Juxta-articular
Nodules,* and they have not, so far as I am aware, been reported
from this part of Africa before.
Of the 964 men, 14 were actually suffering from, and 160 showed
clear indications of having had, yaws: of the 1,414 women, 29 and
234 respectively: for the 567 children, the figures were 15 and 70.
With eight exceptions, the 80 subjects of these subcutaneous
nodules had characteristic scars of, and admitted having had,
yaws: of the eight exceptions, seven had the typical pigmented,
finely wrinkled scars, but denied infection. Yaws is so commonly
Subcutaneous Juxta-articular Nodule on the forearm of a man who had yaws as a child.
contracted by these people in infancy and early childhood that many
may forget all about it by the time they reach adult years. The
opinion which I hold that these subcutaneous nodules are a late
manifestation of yaws, comparable to, but without the tendency to
softening of syphilitic gummata, is not, I think, invalidated by the
above-mentioned exceptions. In most cases the nodules appear
years after the manifest lesions of yaws have healed, but in
four cases they were found in persons still showing unhealed
granulomata.
For comparison, I was able subsequently to examine 327 adults
and 266 children in another (Mlanje) district of the Protectorate
where yaws is not, so far as I am aware, known. The numbers are,
unfortunately, small. No recent case was found amongst these
people, who belong to quite a different tribe from the inhabitants of
Dedza district, and many of whom have, within recent years,
immigrated from Portuguese territory. Only two of the adults
admitted having had yaws or showed indications of infection: both
of these had characteristic yaws scars and also subcutaneous
nodules: one had also beautifully pitted hands and feet—a
condition to which Castellani (1910) has drawn attention. These
two persons had contracted the disease in another part of the
country. None of the 266 children examined showed any indica-
• tions of the disease.
In the Journal of the London School of Tropical Medicine
(1913), in a review of a paper by Ouzilleau, it is stated that in
Mbomau adult Filaria volvulus were found ‘ in cysts or tumours,
generally superficial, situated under the skin,* and ‘ the conclusion
must be drawn that the volvulus cysts situated near joints do not
differ markedly from those tumours known as “ Juxta-articular
Nodules .” 9 ‘ Brumpt had observed many cases of these nodosities
in Uganda and some on the Ouelle * ... ‘ in every case of this
sort seen by the author he had punctured the tumour and found the
embryos and sometimes fragments of the adult of Filaria volvulus 9
I therefore punctured one nodule, and excised two from the forearm
in another case: neither in the small quantity of material removed
by puncture, nor in scrapings from the cut surface of the excised
nodules was anything resembling a filaria seen. Microscopically,
fragments of fibrous connective tissue but no parasites or organisms
of any sort were found. On section the nodules were solid, tough
and lay embedded in subcutaneous fat.
Under the tertiary stage of yaws, Castellani and Chalmers
mention the occurrence of gummatous-like nodules which soften and
break down: such a condition I have observed in a few cases of
yaws: the nodules to which I refer have no such tendency to
softening.
The observations recorded above point strongly, in my opinion,
to these nodules being a late manifestation of yaws, and they so
closely resemble the 1 Juxta-articular Nodules * described in
Castellani and Chalmers' Manual of Tropical Medicine that there
can be little doubt of their identity with the latter.
Since these observations were made, I have read in the Tropical
Diseases Bulletin (1913) that Mouchet and Dubois have observed
similar nodules, and state that the natives in the Congo consider
them a late manifestation of yaws. The sectional editor adds that
Jeanselme's Juxta-articular Nodules are due to a fungus—
Nocardia carougeaui , Brumpt, 1910—but that in Africa, nodules
induced by Filaria volvulus may closely simulate them.
REFERENCES
Castellani and Chalmers (1910). Manual of Tropical Medicine , lit edition, p. 1137.
-(1910). Idem p. 868.
Journal of London School of Tropical Medicine (1913), Vol. II, part 2, p. 133.
Tropical Diseases Bulletin (1913), Vol. I, No. 9, p. 530.
425
ANKYLOSTOMIASIS IN DOGS IN
SIERRA LEONE
\Being the Fifth Report of the Thirty-second. Expedition of the
Liverpool School of Tropical Medicine , 1914-1915.]
BY
WARRINGTON YORKE
AND
B. BLACKLOCK
(Received for publication 9 July , 1915)
Ankylostome infection appears to be universal in dogs in
Freetown; of seven examined by us, all were found to be heavily
infected. More detailed examination showed that the infection was
due to two parasites— Ankylostoma caninum , Ercolani, 1859, and
Ankylostoma ceylanicum, Looss, 1911—which were present in the
intestines in about equal numbers. These' species are readily
distinguished one from the other by the characteristic arrangement
of their teeth. The mouth of Ankylostoma caninum is armed with
three pairs of prominent ventral teeth, whereas in Ankylostoma
ceylanicum there is one pair of large ventral teeth, and one very
small pair near the base of the former, but on a slightly deeper
plane. These characters are illustrated in Figs. 1 and 2.
Fig. 1. Mouth of Ankylostoma caninum.
X 300.
Fig. 2. Mouth of Ankylostoma ceylanicum.
X 300.
426
The bursa of the males is very similar in the two species. It
consists of two large lateral and a small dorsal lobe. The arrange¬
ment of the rays is as follows: In each lateral lobe there is an
anterior ray which is cleft, an antero-extemal ray, a median ray
which is doubled, and a postero-extemal ray which arises from a
common trunk with the single posterior ray. In the dorsal lobe is the
posterior ray, which exhibits slight differences in the two species.
In both it is bifurcated in its terminal third, and each of the branches
is at its extremity tridigitate. It is in the character of these terminal
digitations that the slight difference is found (see figs. 3 and 4). In
Fio. 3. Posterior ray of bursa of Ankylostoma Fig. 4. Posterior ray of bursa of Ankylostoma
caninum . x 500. ceylanicum. x 300.
both species the two inner digitations are small, being separated by
a mere notch. In Ankylostoma caninum , the cleft sparating the
two inner from the outer digits is shallow, but in Ankylostoma
ceylanicum , the cleft is deep, being about half the length of the
branch of the posterior ray.
The average length of Ankylostoma caninum is greater than that
of Ankylostoma ceylanicum. A few were measured by us with the
following results:
Ankylostoma caninum : Males 6*5-8 mm., females 7*5-14*5 mm.
Ankylostoma ceylanicum : Males 6 mm., females 7-10 mm.
The discovery of Ankylostoma ceylanicum in the dog in Sierra
Leone is of interest. This species was first described by Looss in
1911, from a civet cat from Colombo. It was subsequently found
by Lane (1913) in the dog and cat in Bengal, and also in a lion from
the Calcutta Zoological Garden. In the same paper Lane described
this parasite as occurring occasionally in human beings.
Gomes de Faria, in Brazil (1910), found dogs and cats infected
with Ankylostoma caninum and a parasite which he described as a
4 2 7
new species under the name Ankylostoma braziliense 9 but Leiper
(1913)* from a comparison of figures published by Lane and
de Faria, concludes that Ankylostoma braziliense and Ankylostoma
ceylanicum are identical.
We had no opportunity of determining whether Ankylostoma
ceylanicum occurs in human beings in Sierra Leone, where infection
with Ankylostominae is exceedingly common. The importance of
ascertaining whether this species occurs in man is obvious. If
Ankylostoma ceylanicum is found in human beings in Freetown, the
dog reservoir of the infection is a factor which must be borne in
mind when prophylactic measures are under consideration.
CONCLUSIONS
Dogs in Freetown are heavily infected with Ankylostominae.
The species found were Ankylostoma caninum and Ankylostoma
ceylanicum; they were present in about equal numbers.
REFERENCES
de Faria, G. (1910). Contribution towards the classification of Brazilian Entozoa. Memorial do
Instituto O steal do Cruz , II, pp. 286-293.
Lank, Clayton (1913). Tricbostrongylus columbriformis (Giles, 1892), a Human Parasite. Ind.
Med. Gaz ., XLVIII, pp. 129-132.
-(1913). Agcbylostoma ceylanicum , a New Human Parasite. Ind. Med. Gaz ., XLVIII,
pp. 217-218.
Lkipxr, R. T. (1913). The Apparent Identity of Agcbylostoma ceylanicum (Looss, 1911) and
Agcbylostoma braziliense (Faria, 1910). Journ. Trop. Med. & //yg., XVI, pp. 334-335.
Looss, A. (1911). Records of the School of Medicine , Cairo, Vol. IV.
4 2 9
STUDIES IN BLACKWATER FEVER*
IV.—NOTE ON A CASE OF QUARTAN
MALARIA ASSOCIATED WITH
BLACKWATER FEVER
BY
J. W. W. STEPHENS
SIR ALFRED JONES PROFESSOR OF TROPICAL MEDICINE, THE UNIVERSITY OF LIVERPOOL
(Received for publication 2 July , 1915 )
One Chart
Past History
X.Y.Z. European, W. Africa. Length of residence in Africa
four tours, t
31.3.13. —6.4.13. Sunstroke.
29.4.13. — 3 . 5 . 13 . Sciatica and malaria. Has been ill with
more or less fever for last seven days
Present History
12.8.13. 4 p.m. Admitted to hospital. Condition : Skin yellow,
unhealthy looking; conjunctivae pale; gums pale; tongue dry
yellow-brown fur. Liver palpable i^* below costa. Pulse, 108,
Temperature, ior8°.
14.8.13. Patient progressing favourably until
15.8.13. at 4 p.m., feeling of distress, anorexia, frequent shivers,
inclination to vomit. 10.30 p.m. Blackwater passed.
16-19.8.13. Frequent vomiting, extreme jaundice, increasing
heart-failure.
18.8.13. Slight oozing of blood from gums.
19.8.13. Profound collapse. Death 1.20 p.m.
• Part I j Annals of Trop. Med. & Parasitol 1913. Part II: Ibid., 1914.
Part III : Ibid., 1915, p. 201.
f A tour is a year.
43 °
In the accompanying tables I have given an abstract of the
available data as to the case. They are presented here for the sake
of completeness rather than for any bearing they have on the
problem raised.
Record of Quinine
12.8.13., 4 P* m * Quinine grs. vi. intramuscularly.
13.8.13. ‘Mist, quin.' quartis horis (at what hours given not
stated).
14.8.13. Presumably ‘Mist, quin.' continued.
15.8.13. Presumably continued during the day, ? till 4 p.m..
as it is stated that in the evening it was discontinued, but whether
before or after the blackwater is not stated. Aspirin 10 grs.
presumably at or after 4 p.m.
(NOTE. —The Mist, quininae of the B.P.C. contains gr. i. to 31,
but whether the above mixture is this, it is impossible to say.)
Blood
Red cells
Hgb.
Total
count
leucocytes
Percent¬
age
count
number
counted
Urge
mono¬
nuclear
Lympho¬
cytes
Poly¬
nuclear
Eosino¬
phil
Parasites*
12.8.13 •••
Quartan parasites,
13.8.13 ...
...
...
...
...
...
...
...
scanty rings
14.8.13 ...
...
...
•••
...
...
15.8.13 ...
m.)
2,500,000
...
300
6
it
83
Parasites negative
(10.30 p.
16.8.13 ...;
5 °%
(Tallqvist)
...
...
...
...
1
17.8.13 ...
1,400,000
3 °%
-
...
'
...
18.8.13 ...
6oo,ooof
2 5 %t
18,000
300
H
15
65
6
Parasites negative
* Five Blood examinations were made, but the dates of two of them are not stated,
t Non.—These data imply a color index of 2*i 1
43i
UrIKE
Date
Time
Quantity
Reaction
Colour
Hgb.
Casts, etc.
.
...
...
...
No alb.
Bile present
14.S.13 .
1 5 - 8 -'3 .
Apparently
... 1 ... 1
no urine passed before 10.30 p.m.
5 ) .
10.30 p.m.
220 c.c.
Faintly alkaline
Black
Met. Hgb.
Granular casts
16.8.13 .
2 a.m.
70 c.c.
Faintly add
»»
ss
» .
3 a.m.
14 c.c.
Add
Rather lighter
yy
IS .
9 a.m.
145 c.c.
Aik.
...
ss
ss
n .
9.40 a.m.
170 c.c.
(In the motion)
...
>s .
12.15 p.m.
240 c.c.
Aik.
Met. Hgb.
yy
» .
6 p.m.
240 c.c.
ss
Much lighter
yy
yy
17.8.13 .
12.30 p.m.
278 c.c.
Add
Darker
yy
No casts
» .
—
230 c.c.
Aik.
»
yy
yy
18.8.13 .
2 a.m.
360 c.c.
Acid
Clearing
yy
yy
ss .
11.45 a.m.
50 c.c.
ss
Amber
yy
■
yy
ss .
10.30 p.m.
155 c.c.
ss
Light amber
No alb.
Trace of bile
» .
11 p.m.
20 C.C.
ss
n
yy
yy
1
19.8.13 .
12.45 a * m *
60 c.c.
yy
yy
yy
,
yy
ss .
12 noon
?
ss
...
...
No casts
Post-mortem (partial) Fite Hours apter Death
Skin .
Very yellow
Liver .
Very large, yellow mottled,
Fatty degeneration, perilobular
soft, old adhesions
fibrosis. No parasites, some
pigment
Spleen .
Large
No parasites, some pigment
Stomach.
Petechiae oozing blood, con¬
tents green bile
Small intestine.
Petechiae oozing Mood
Gall bladder .
Yellow bile
j
Kidneys .
Large, pale
Many lobules blocked with
‘ haemoglobin ’ casts
Bone marrow .
Pale
No parasites, some pigment
+ 3 *
This case is of interest for the following reasons.
1. Immediately after the blackwater, 15.8.13, 10.30 p.m., no
malaria parasites or pigment were found, i.e., there is no evidence
of a malaria infection, yet on 12.8.13, 4 P-m., scanty quartan
parasites were found, i.e., the patient at that time was infected with
malaria. The records do not permit of any conclusion as to when
evidence of malaria infection ceased, for although two other blood
examinations (presumably negative) were made, yet it is not stated
when they were made, i.e., whether before or after the blackwater.
Further, the only evidence of a malaria infection post-mortem is the
presence of pigment, parasites being absent. It is improbable,
however, that a quartan infection was got rid of by three days’
quinine treatment. We have then a case of blackwater fever,
negative as regards malaria, subsequent to the blackwater, but
positive three days previously, and also positive (pigment) post¬
mortem.
2. The diagnosis of quartan parasites on the 12th is supported
by the character of the chart (see p. 433), which is quartan in
character. If we assume that the rise on the 15th is a quartan
paroxysm, then we have the actual paroxysm in this case, for some
reason or other, associated with blackwater. It may be argued,
however, that the paroxysm on the 15th is due to the onset of black¬
water per se y and that it is a coincidence that it has occurred when
we should have naturally expected a quartan paroxysm. This
possibility cannot be disproved, but also the possibility—if nothing
more—exists that this is not so, and that in this case a quartan
paroxysm is closely concerned with the blackwater attack. The
chances appear to me to favour this interpretation, because the
quartan paroxysm would occur on this day, whereas the blackwater
might have occurred at any other time. A study of temperature
charts (unfortunately few are available), previous to the onset of
blackwater, would probably throw light on this point.
I hope, in due time, to be able to collect a number of suitable
cases. I have to acknowledge with pleasure the kindness of the
Colonial Office in putting the records of this case at my disposal.
Volume IX
December, 1915
No. 4
ANNALS
OF
TROPICAL MEDICINE AND
PARASITOLOGY
ISSUED BY
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE
Edited by
Professor J. W. W. STEPHENS, M.D. Cantab., D.P.H.
Professor R. NEWSTEAD, M.Sc., J.P., F.R.S., A.L.S., F.E.S., Hon. F.R.H.S.
Professor WARRINGTON YORKE, M.D.
AND
Professor Sir RONALD ROSS, K.C.B., F.R.S., M.D., F.R.C.S.,
Major I.M.S. (Ret.)
Editorial Secretary
Dr. H. B. FANTHAM,
School of Tropical Medicine ,
The University ,
Liverpool.
C. 'Tinling & Co. t Ltd.
Printers to the University Press of Liverpool
$3 Victoria Street
435
NUCLEAR VARIATIONS OF THE
NEUTROPHILE LEUCOCYTES
(ARNETH COUNTS) IN MALARIA
AND YELLOW FEVER
BY
J. W. SCOTT MACFIE, D.Sc., M.B.
WEST AFRICAN MEDICAL STAFF, GOLD COAST
(Received for publication 22 July , 1915)
Considerable interest has recently been evinced in the changes
observed to take place in various diseases in the nucleus of the
polymorphonuclear leucocyte; and, following Ameth’s method of
classification of these cells, it has been shown that there is an increase
of the cells of Classes I and II in many microbic diseases. The
researches of Chamberlain in 1910 first drew attention to the fact
that a similar phenomenon is exhibited by persons living in the
tropics; and it has recently been suggested by Breinl and Priestley
(1914), as the result of their examinations of school children in
Queensland, that this change may be ‘ due to the effects of a tropical
climate upon the white race living in the tropics/
An increase of the percentage of polymorphonuclear leucocytes
belonging to Classes I and II has been observed in a great many
different diseases. It occurs, according to Cooke (1915), in
typhoid, scarlet fever, diphtheria, measles, chicken-pox, erysipelas,
tonsillitis, pneumonia, whooping-cough, puerperal fever, tuber¬
culosis, nasal catarrh, and septic conditions, and probably in other
diseases also. It is important, therefore, in considering the
significance of a shift to the left of the Arneth count, such as that
found in Europeans in the tropics, to determine in the first instance
the effect on the polymorphonuclear leucocytes of the diseases
prevalent in these climates, and especially that of such an infection
43 ^
as malaria which may lie latent in the body for long periods without
exciting active symptoms. In the following pages are embodied the
results of the examinations of a number of cases of malarial fever
and yellow fever with special reference to the changes occurring in
the polymorphonuclear leucocytes.
The blood-films were all stained by Leishman's method. Two
hundred successive neutrophile leucocytes were examined in each
film, each hundred being counted separately and the two sets of
figures being accepted only if they agreed approximately. Nuclear
fragments connected by a fine thread were counted as separate; but
those joined by a definite band were counted as one. In all
doubtful cases the cells were referred to the higher of the two classes
to which they might have been assigned.
In making Arneth counts there must always be a very consider¬
able personal variation. Many cells are exceedingly difficult to
assign to any particular class, and in such cases different investigators
might easily differ in their ultimate decisions. The various
estimates of the normal Arneth index, which range from twenty-five
to fifty-six, are in themselves a proof of this fact. Allowance must
always be made, therefore, for some degree of individual difference
in comparing counts made by different observers, and too great
importance should not be attached to small degrees of shift. A
series of counts by the same observer in different conditions should,
however, be strictly comparable. In making the counts referred to
below, great pains were taken to try and follow the course of the
nucleus before attempting to decide to which class the cell should
be assigned. It was assumed that the nucleus was a continuous
body with two free ends, and that it should be possible to trace the
connexions between the different lobes. This assumption was not
always borne out in fact, as leucocytes are occasionally met with
even in normal blood in which the nuclear fragments are actually
separate from one another. Such cells are probably dead cells; but
as they are invariably easy to classify, they do not affect the counts.
In my experience the greatest difficulties occurred in films which
showed a normal, or nearly normal, count. In those in which there
was a marked shift to the left, and these were the most interesting
of the series, there was seldom any doubt whatever as to the classes
in which the various leucocytes should be placed.
I. THE RESULTS OF ARNETH COUNTS IN HEALTHY
EUROPEANS AND NATIVES
A. Europeans.
Twenty-nine apparently healthy Europeans were examined at
various stages of their tours of service in West Africa (see Table I).
The average Ameth index was 51 *6; that is, there was a slight shift
to the left in the count of about the same degree as that found by
Chamberlain and Vedder (average index 46*2) in seventy-two healthy
American soldiers in the Philippine service.
Eleven of the subjects had been in residence in the Gold Coast
for less than a year, and eighteen for twelve months or longer. The
Ameth index in the former averaged 50 6, and in the latter 516;
a scarcely perceptible increase in favour of those who had been in
West Africa for the longer period.
Two of the Europeans had landed at Accra only one and four
days respectively before they were examined. In the latter the
Ameth count was practically normal (index 41*5), but in the former
there was a slight but distinct shift to the left of the count for which
there was no apparent cause. He had previously served in West
Africa, and had, of course, been in the tropics during the last few
days of the voyage before he landed, and had possibly, although
not probably, been infected by mosquitos. But in this connexion
it should be noted that it is not a very uncommon occurrence for men
returning to West Africa to show symptoms of malaria soon after
landing; and that this is probably due to their having failed, during
their leave of absence, to eradicate from their systems the malarial
infections contracted during their previous tour of service.
In a subsequent part of this paper it is shown that in malaria
there is a pronounced shift to the left of the Ameth count, and that
this phenomenon is observable before the parasites are sufficiently
numerous to be found in the peripheral blood, and that it may
persist for a considerable time after the patient has apparently
recovered. Practically all the apparently healthy Europeans
examined had suffered from malarial fever at some time during their
tour, and it may be assumed that they had all been repeatedly
inoculated with malaria parasites by infected mosquitos. I believe
that this is a sufficient explanation of the slight shift to the left of
Arneth classification per cent.
No. Sex Length of tour __Arneth
I
11
III
IV
V
Index
1
M.
4 days
5*5
36-0
41-0
* 5*5
2*0
4**5
2
M.
1
day
8-5
45-0
39 -o
7*o
o*5
53*5
3
F.
3 months...
6-o
3 °*°
47*5
14*0
2 *5
36-0
4
M.
6
I o-o
29*0
37 *o
21*0
3 *o
39 *o
5
M.
6
« -
5 -o
41*0
39*5
* 3*0
**5
46-0
r>
F.
7
,2 *5
38-5
34*5
,2 *5
2*0
51-0
7
M.
10
1 ?
9*5
4**5
37*5
11*0
0-5
51*0
8
M.
10
11
* 3*5
47 -o
30*0
9*0
o*5
60*5
9
F.
10
„ ...
16-o
47 -o
2 9*5
7*5
—
63*0
IO
M.
11
,, ...
17-0
38*5
36*5
8-o
—
55*5
11
M.
11
V
* 3*5
46-5
36-0
4 -o
—
6o*o
12
M.
12
11
15-0
3 8-8
35*o
10*0
1*2
53*8
13
M.
12
11
22-0
46*0
29-0
3*0
—
68*0
14
M.
12
11
130
45 *°
37*5
4*5
—
58-0
*5
M.
12
T)
17-0
40*0
35*o
8 -o
—
57 *o
16
M.
12
11
7-5
4**5
36*0
14-0
1*0
49-0
*7
F.
12
11
4-0
41-0
42-0
130
—
45 -o
18
M.
12
11
i 10*0
4 2 *5
37*o
9*5
1*0
| 5 2 ‘5
*9
F.
12
5J
j 16*0
j
39*5
33*5
11*0
55*5
20
M.
13
?)
18-0
37*o
33*5
IO '5
1 *0
55 -o
21
M.
13
n
7*5
1
44.0
40*0
8 -o
o*5
5**5
22
M.
13
M
18-0
57 *o
2 3’5
*•5
—
75 *°
2 3
F.
13
»
9*5
43*5
37*5
9*5 :
—
53 -o
2 4
M.
13
135
41*0
36*0
9 *o 1
t
0-5
54*5
2 5
M.
14
J?
7 *o 1
33*o
41*0
i 7 *o |
2*0
40*0
26
M.
14
V
8 -o
38*5
4 2 *5
10*0 '
1*0
46-5
2 7
M.
»5
11
5*5
35 *o
43 *o
1
14,5
2*0
40*5
28
M.
15
1)
8 -o
2 4*5
40*0
i
2 3*5
4*0
3 2 ’5
2 9
M.
16
11
15-0 |
37 *o
35*o
IPO
2*0
52*0
Averages
|
u 4 ;
1
40*2
367
107
1*0
51-6
439
the Ameth count found in these subjects. I do not consider that
there is at present sufficient evidence in support of the view that this
phenomenon is due to the influence of a tropical climate on the white
race. The fact that the shift to the left was almost the same in those
that had been in West Africa over a year, and in those who had but
recently arrived, is, I think, contrary to any such supposition.
It is clear, at any rate, from these examinations that certain
individuals may show a normal Arneth count after more than a year’s
residence in West Africa (see Table I, cases Nos. 25 and 26). Some
of the individuals in whom the count approximated most nearly to
the normal were men who had spent many years on the 4 Coast,’ and
who might be supposed to have become acclimatised, or to have
mastered the conditions of life most suitable for West Africa. Others
were individuals who were on the whole better cared for than the
majority of their fellows, who had better quarters, and who were to
a great extent relieved from the petty domestic worries that are
responsible for so much irritation, and probably so many of the
ailments, in West Africa.
B. Natives.
Ameth counts were made in the cases of twenty apparently
healthy natives at Accra. All the blood films were taken on the
same day, and within the same hour, from twenty labourers who
were actually at work at the time when they were called up for
examination. The average index worked out at 55*92, and the
averages of the percentages of the polymorphonuclear leucocytes
belonging to the five classes of Arneth’s classification showed a slight
shift to the left (see Table II).
There were, however, great variations in the different counts;
and the index ranged from 26'5 to 98*5. It is doubtful, therefore,
if an average based on such a small number of counts is of any value.
A better idea of the actual conditions found is obtained, I believe,
by distributing the indices into groups, and plotting a curve to show
the percentages of the cases that fall into each group. This has
been done in Chart I. It will be observed that the crest of the
curve in natives shows a slighter degree of shift to the left from the
normal point than that in Europeans; although in the former the
average index worked out at 55*92, and in the latter at 51‘6.
440
Table II.—Arneth counts in apparently healthy natives at Accra, West Africa.
1
No. ,
Arneth classification per cent.
1
Arneth
index
i
I
II
III
, IV
V
I
7.0
44*5
00
6
, 9*5 1
1*0
; 51*5
2
10*0
42-0
41*0
7-5 i
°*5
! 5 1- °
3
24*0
52*0
22*0
2*0 j
—
1 76*0
4
22*5
51-0
24*0
2,5 !
—
73*5
5
95
37*5
39*5
12-5 !
1*0
47 -o
6
] ' 5 '°
36-0
39*5
9’5
—
51-0
7
2 ‘5
24*0
! 48*0
2 1 *5
4 *°
26-5
8
10*0
37 *°
1
4 °'°
* 3 °
—
47 -o
9
13*0
37 '°
1 37'5
12-5
—
50*0
10
8 0*5
i8*o
I'O
—
°’ 5 #
98-5
11
12-5
36*5
00
6
1 I'O
2-0
49.0
12
10*0
36-5
38-5
130
2-0
46'5
n
8-o
34 ’°
l
40*0
17*o
1*0
42*0
14
16*o
4 I *5
33*5
8-o
1*0
57*5
*5
* 4-5
49-0
i 3°*5
6 *o
1 -
63-5
16
i 8‘5
48*0
29-0
4*5
1
66-5
17
! 4'5
35*5
00
Wl
I 1*0
1
°*5
i 50*0
18
i o *5
35*0
43-5
105
°'5
1 45*5
!
19
! 3-5
36-5
39-5
9*5
1*0
i s°*°
20
3**5
43-5
23-0
2*0
—
j 75 *°
17-17
3*75
1 34 * 22
9-15
075
! 55 ' 9 Z
• This leucocyte was probably a dead one undergoing chromolysis. I am unable to
account for the very high index in this case. The man when seen again a day or two later
appeared to be in good health, and no information could be elicited excepting that he had
had 4 fever a short time ago/ His Arneth index four days later was 64-5.
The shift to the left of the Arneth count in natives at Accra was
only a slight one. It was much less pronounced than that discovered
by Chamberlain and Vedder in Filipinos (average index 65*8).
There are several factors in the normal life of healthy natives
that may have a bearing on the condition of the blood, especially
the prevalence of malarial and intestinal infections.
Chart I.—The distribution by groups of the Arneth index in (A) twenty
healthy natives, and (B) twenty-nine healthy Europeans at Accra.
It may be assumed that nearly every native harbours intestinal
parasites, since ova of various species of worms, Trichomonas , and
spirochaetes are very generally found in the faeces, and adult worms
are recovered at practically every post-mortem examination. Of the
blood changes produced by these infections anaemia and eosino-
philia are the best known; but Knapp (1915) has recorded that he
has found a shift to the right of the Arneth count in amoebic
dysentery and ankylostomiasis.
With regard to malaria, plasmodia are to be found in the blood
of all, or almost all, native children; and every native in the course
of his normal daily life must be repeatedly inoculated by infected
mosquitos. I believe that in natives, as in Europeans, the shift to
the left of the Arneth count found in apparently healthy individuals
may in most cases be accounted for by such malarial infections.
Other diseases may, of course, be responsible in some cases; such,
for instance, as tuberculosis, or the mild attacks of yellow fever that
scarcely incapacitate the natives at all. But as malarial infections
dominate the field in tropical medicine to so great an extent, and
as in this disease there is so pronounced a shift to the left of the
Arneth count, it is not unreasonable to suppose that in the majority
of cases the shift in apparently healthy individuals is due to this
cause.
If infections with intestinal parasites (amoebae, worms,
Trichomonas , spirochaetes, &c.) are proved to cause a shift to the
right of the Arneth count, there must be a condition of balance in the
cases of most natives in which the shift to the left resulting from
malarial infections on the one scale weighs against the shift to the
right due to intestinal parasites on the other. And the same
condition may occur in latent infections with syphilis and leprosy,
diseases in which Knapp also noted a shift to the right.
II. THE RESULTS OF ARNETH COUNTS IN MALARIAL FEVER
In a previous note (1915) it was pointed out that in malarial
fever there is a well marked shift to the left of the Arneth count, and
that this phenomenon persists after all parasites have vanished from
the blood, and after the patients have recovered from the attacks.
At the time this preliminary note was published I had not seen any
other recorded observations on the nuclear variations of the poly¬
morphonuclear leucocytes in malarial fever. Recently, however,
I have received a copy of the Indian Medical Gazette (March, 1915)
containing a paper by Knapp on ‘ The Significance of Arneth’s
Leucocyte Count,’ in which the author records that he has found a
JXtcv>r<\joy>VvJjL £4^coc^Vn sitJtow^cv^ h) CoCK ctaoo.
443
Chart II.—The average Arneth count in (N) twenty healthy natives,
and (E) twenty-nine healthy Europeans, at Accra, compared with
the normal count in Europe (Arneth)
444
decided shift to the left in malaria. Knapp states that 4 Out of
21 cases, it was marked in 4, definite in 13, and absent in only 4/
that it was, on an average, 4 equal in tertian and subtertian fevers,
and a single case of quartan gave a similar finding 1 ; and he
concludes that 4 on the whole it may be fairly said that absence of
the left-shift is presumptive evidence against malaria.’ Particulars
of the counts are given in only two cases: the one, a case of 4 Double
Tertian,’ had an index of (?) 669, and the other, 4 Subtertian,’ an
index of 59*4.
During the last few months I have performed Arneth counts in
twenty-three cases of malaria in which the diagnosis was confirmed
by the finding of the parasites, nine cases of fever in which there was
presumptive evidence of malaria, namely an increase of the large
mononuclear and transitional leucocytes to 15 per cent, or over, and
twelve cases of fever suspected of being malarial but in which neither
positive nor presumptive evidence of infection could be found.
That is, counts have been made in forty-four cases of proved or
suspected malarial fever.
Of the twenty-three cases in which malaria parasites were found,
ten were Europeans, one a Syrian, one a half-caste child, and eleven
natives. In each case a very marked shift to the left of the Arneth
count was found (see Table III). My results were, therefore, much
more uniform than those of Knapp, and the shift observed was much
more pronounced. In the Europeans the index averaged 84 25, and
in two cases was as high as 95*5. In the natives it was even higher;
the average being 897, and the highest 97*5. P. falciparum was the
only species of parasite present in nineteen of the cases, and
P. malariae in two. One case showed both P. falciparum and
P. malariae , and one P . falciparum and P. vivax (see Table III).
A rough estimation of the number of parasites present in the
peripheral blood was made in each case, and the results are shown
in the Table as a ratio of the number of red corpuscles to each
parasite. An examination of the figures shows that there is no
direct relationship between the number of parasites present in the
peripheral blood and the degree of deflexion to the left of the Arneth
count. The deflexion was greatest in those cases in which the
constitutional disturbances were greatest; and, in the case of natives,
in children.
445
Table III.—Ameth counts in twenty-three cases of malarial fever.
No.
Race
:
Arneth classification per cent.
Nature of the infection !
Ameth
I
!
i
n
III
IV
V
1 index
i
European ...
P. falciparum , 1:1-4 R*B.C. A few P. 78*0
malariae. Fatal case
I 7'5
4-0
o -5
—
95*5
2
11
P. falciparum , 1 : 700 R.B.C. .51*0
I 3 2 ’°
16*o
PO
—■
00
U 4
6
3
11
P. falciparum , 1 : 625 R.B.C. .22*0
53 *o
23-0
2*0
—
75 -°
4
11
P. falciparum , 1 : 250 R.B.C. .27*0
47.0
1
24-0
2*0
—
74 -o
5
11
P. falciparum , 1 : 700 R.B.C. .27*5
5°*5
* 9*5
2 '5
—
'vj
00
6
6
11
P. falciparum , very rare. Blackwater fever. 6o*o
Fatal
27*0
1
12*0
PO
—
87-0
7
11
P. falciparum , 1 : 714 R.B.C. .56*5
39-0
4 -o
°*5
—
95*5
8
11
P. falciparum , 1 : 830 R.B.C. .43-5
46-5
10*0
—
—
90-0
9
»>
P. falciparum , few. Blackwater fever, first 29*5
day. Mild case
45-0
2, ‘5
4 -o
—
74*5
IO
11
P. falciparum , 1 : 8500 R.B.C. . 41*0
49*0
90
PO
—
90-0
"
Syrian
P. falciparum , 1 : 1250 R.B.C. . 24*0
56'0
17*o
3 -o
—
8o-o
12
Half-caste ...
P. falciparum , 1 : 20 R.B.C . 30*0
53-5
16-o
o -5
—
83*5
>3
Native
P. falciparum, 1 : 400 R.B.C. . 67*0
! 2 5 *° 1
8-o
—
—
92*0
11
P. falciparum, 1 : 713 R.B.C. . 46*0
1 43 *°
II-O
—
—
89*0
>5
»
P. malariae, very rare .j 24*0
5 2 *5
21*0
2 * 5
—
76-5
16 i
11 •••
P. malariae , scanty . , 57*0
34 *o |
9 *o
—
—
91*0
17
,,
1
P. falciparum (crescents only), 1 : 5000 ; and 32*0
P. vivax, 1 : 4100 R.B.C.
1
55 *°
* 3 *°
— 1
—
8 7 -o
iS j
??
P. falciparum, 1 : 100 R.B.C. .35-0
48-0 j
16*o
PO ,
—
83-0
19
11
P. falciparum, 1 : 19 R.B.C. 1 54*5
35 *o
10*0
-5;
—
89-5
20 ,
11
P. falciparum, very few .56-0
4 i -5
2 ’5
1
—
■ 97-5
21
11 •••
P. falciparum, scanty . 39*5
53 *o
7*5
— 1
—
9 2 ’5
22
11
P. falciparum, very rare .64*0
33 *o ,
3 -o
1
i 97 *°
23
11
P. falciparum, 1:91 R.B.C.54-0
37*5
7*5
PO |
—
9 p 5
Averages of the counts in the twenty-three cases . 1 44*3
4 2 *3
12-4
PO |
_!
—
86-6
446
The shift to the left was also well marked, but less in degree, in
those cases of fever in which there was only presumptive evidence of
malarial infections (see Table IV). The index averaged 79*3 in the
Table IV.—Ameth counts in cases in which no malaria parasites could be found, but in which
the large mononuclear and transitional leucocytes numbered 15 per cent, or over.
Arneth classification
per cent.
Differential counts
per cent.
Race
Arneth
0
-5 u
S
>.
o’Sjs !
g S 2 i
eft
J V
I
II
1
III
,v
V
index
Polymorp
nuclea
1
s
J
= S;=
&|Sj
1
_c
OU
0
c
•a
0
w
0
u
rt
%
European
59 -o
36*5
4‘5
—
—
95*5
74 -o
IPO
* 5 *° 1
—
M
39*5
47-0
I 2*0
'•5
8rt- 5
6P5
20*5
, 6 - 5 ;
PO
o -5
33
3 i *5
50*0
17*0
i *5
—
81-5
5PG
31*5
16*5 1
PO
—
51 •••
34 *o
50*0
15-0
1*0
—
84-0
4 2 "5
37*5
i8-o ,
p 5
o -5
,, ...
8-o
5°*5
| 3 »*° 1
10*0
°*5
58-5
56*0
22*5
.8-5
2-0
PO
33 •••
24-0
3^*5
l 28-5
9’5
p 5
60-5
54 -o
l 4 -0
!
20*0 |
12*0
—
” "1
33 -o
49 *°
1
i To
1*0
| 1
1
82*0
66-o
12*0
2I *5 1
1
o -5
—
33
33*5
46-0
j T 5
3 -°
79*5
59*5
6*o
34*5 ;
—
1 -
Native
40-0
46*0
1
I 3 ’° ,
PO 1
1
I ;
86*o
59 -o
25-0
1 5 '° 1
o -5
!
! 0-5
Averages
33-6
457
1 T 3 '
1 1
3-2 i
0*2
79*3
58-2
20-0
1 9*5
I
2*0
i °-3
nine cases, and ranged from 58*5 to 95 5. No direct connexion
between the percentage of the large mononuclear leucocytes and the
degree of the shift could be traced. This was not to be expected,
as the blood films were taken at different stages of the fever, and it
is well known that the percentage of large mononuclear cells is
subject to great fluctuations in the course of the disease.
Of the other twelve cases of fever, seven were in Europeans, and
five in natives. The seven cases in Europeans were all diagnosed
as malarial from their clinical appearances, but no proof of infection
could be obtained by the examination of the blood. In West Africa,
where practically every European takes five grains of quinine daily,
it is often extremely difficult to find evidences of malarial infections
in the blood in cases of fever which look like malaria, and which
447
respond favourably to treatment with somewhat larger doses of
quinine. There can be little doubt that the majority of such cases
are actually malarial; but they are a source of difficulty and anxiety
to the physician, and any sign that would assist in the diagnosis
would be of great value. It is interesting, therefore, to note that in
all these seven cases there was a decided shift to the left of the
Ameth count. The index averaged 71 ‘35; the lowest being 61 5,
and the highest 82 0 (see Table V).
Table V.—Ameth counts In cases of fever suspected of being malaria, but in which neither
positive nor presumptive evidence could be found.
j Ameth classification per cent.
No.
Race
1
II
III
IV
V
Ameth
index
A 1
European
1 7*5
39 -o
32-5
1 o-o
1-0
56-5
2
» .
39 *o
41-0
18-o
2-0
—
8o-o
3
..
i8-o
48-5
29-0
4‘5
—
66*o
4
33 -o
49-0
16-o
2-0
—
82-0
5
» .
15-0
46 -5
29*5
9 -c
—
61-5
6
” .
23-0
5»*5
23-5
2*0
—
74*5
7
30-0
49-0
18*0
3-0
—
79.0
Averages
i
: 25-07
46-36
2379
4-64
0-14
7 * *4
B 1
| Native .
i
j 37-5
1
49*5
12-0
1-0
—
87-0
2
11 ... ... ...
1 4*0
28-0
50-0
15-0
3 *°
32-0
3
.
16-o
; 53 -o
26-0
5-0
—
69-0
4
16*5 |
|
54.0
2 7 -5
2-0
—
70-5
5
» .
7-0 j
44 *o ,
41-0
8-0
—
51-0
]
Averages
16*2
t
457
3**3
6-2
o-6
61-9
In malarial fever, therefore, there is a marked shift to the left of
the Ameth count, which may be extreme in degree in cases accom¬
panied by severe constitutional symptoms, but is still quite definite
448
even in cases in which there is no proof of the nature of the infection
other than the clinical and therapeutic one. In severe cases the
majority of the polymorphonuclear leucocytes are of the types
included in Class I, and forms with simple horseshoe-shaped nuclei
are peculiarly abundant. On examining a blood film from such a
case one is at once struck by the uniform and unfamiliar appearance
of the polymorphonuclear leucocytes, and the almost complete
absence of the familiar normal forms with the nucleus divided into a
number of separate lobes joined together by delicate threads.
This effect on the neutrophile leucocytes is observable before the
onset of the attack of malaria, and before the parasites are
sufficiently numerous to be detected in an examination of the
peripheral blood. An example may perhaps be given. On
April 2 ist I examined the blood of a European who had landed at
Accra three weeks previously. He had recently returned from
leave, and had not taken any quinine for six months, but had
enjoyed good health during all this time. I found that there was a
marked shift to the left of the Ameth count, and that the index was
61*5; and although no parasites and no pigmented leucocytes could
be found, and the percentage of the large mononuclear cells was not
higher than that frequently found in healthy Europeans in West
Africa, I suggested that it was probable that he had been recently
infected with malaria. A few days later he developed a typical
attack of malarial fever.
This shift to the left may persist for a considerable time after the
attack of malaria is apparently cured. In Table VI the results of
five successive Ameth counts on the same individual are shown.
Table VI.—Successive Ameth counts in a severe case of malarial fever in a native.
Date
Clinical condition
Arneth classification per cent.
'
Ameth
index
I
1 II
m
IV |
V
Jan.
14, 1915...
P, falciparum , 1 to 400 R.B.C....
67*0
! 25-0
8-o j
— |
—
92*0
Jan.
15, 1915...
P. falciparum, l to 713 R.B.C....
46*0
43 ’°
11*0
— I
—
89*0
Jan.
25,1915...
Well .
2 7*5
47-0
22*0
3*5
—
74*5
Feb.
9,1915...
Well .
20*0
4*’5
|
34*5
1
3 -o
—
62-5
Feb.
2+, 1915...
Well .
18.5
43*5
34-0
4 -o j
—
62*0
+49
This patient, a native, was treated with quinine only during the
actual attack of fever; and although he made a rapid and apparently
complete recovery, there was still a decided deflexion of the Arneth
count six weeks after the onset of his illness.
In West Africa at the present time repeated infection with the
parasites of malarial fever must be recognised as one of the normal
factors of everyday life alike for the native and the European. The
native by his tolerance, and the European by his prophylactic doses
of quinine, is able, as a rule, to ward off the actual attack of fever;
but the parasites are continually being introduced into the system,
and sooner or later they obtain a foothold. Once infected, it is well
known how difficult it is to eradicate them completely from the body.
Bearing these facts in mind, I think it is only natural that the
average native and European in West Africa should show evidences
of malarial infection, and I believe that the shift to the left of the
Arneth counts found in healthy individuals (see Section I) are to be
explained in this manner.
III. THE RESULTS OF ARNETH COUNTS IN YELLOW FEVER
The early diagnosis of yellow fever is a matter of great difficulty,
especially in the mild attacks which occur in natives in West
Africa. In the Second Report of the Yellow Fever Commission
(West Africa), one of the conclusions arrived at is that * the extreme
practical importance of being able to determine whether a mild case
of fever is or is not yellow fever renders it essential that all possible
methods should continue to be employed in the clinical study of the
disease/ and it is recommended that ‘ the attention of all workers at
this subject should be specially directed to the discovery of a
clinical test for yellow fever/
I was anxious, therefore, to carry out Arneth counts in cases of
yellow fever, since this test, should it prove to be of any assistance,
would be a very simple one for the physician to employ, as it does
not entail any elaborate technique. Unfortunately, the single case
of this disease that has occurred at Accra since my arrival was only
identified at the autopsy. Recently, however, Dr. J. M. O’Brien
has been so kind as to hand over to me a series of blood-films he
had collected from cases of yellow fever at Guayaquil, Ecuador;
and has permitted me to make use of them for the purposes of this
investigation. I wish to take this opportunity of thanking
450
Dr. O'Brien for his generosity in allowing me to make use of this
material.
The slides were from the cases of yellow fever in which
Dr. O’Brien (1914) detected a degeneration of the polymorpho¬
nuclear leucocytes which he believes to be characteristic of the
disease. On examining them I found that they had been taken on
thirty-five days from seventeen different cases of yellow fever. In
seven cases there was only a single blood-film, but in the other ten
there were two or more, taken on successive days of the disease.
Ameth counts have been made on all the thirty-five films, but I
shall limit myself for the present to a consideration of those made
on the first days of observation only, the details of the counts on
which will be found in Table VII.
Table VII.—Ameth counts in yellow fever.
No.
Arneth classification f
>er cent.
1
! Arneth
I
II
III
IV
V
index
1
1
43-0
41*0
1 5*5
1 o -5
—
■
0
rh
00
2
48*0
‘ 40-5
9-5
1 2-0
—
I 88-5
3
67-5
30-0
2*0
°*5
—
97*5
4
69*0
30*0
1*0
—
—
99.0
5
82*0
17*0
1*0
—
—
99 ‘°
6
40*0
40*0
19*0
1*0
—
8o*o
7
49-0
44.0
7 *o
—
—
93 *o
1
8
58-5
36*5
5 -o
—
—
1 95-0
|
9
75 *o
24*0
1*0
!
—
99 *o
10
34-0
50*0
15*0
1*0
—
00
4*
6
11
53 *o
43*0
4 -o
—
i
96-0
12
20*0
5 2 *5
22*0
5 -o
°*5
7 2 -5
>3
55*5
34 *o
8*5
2*0
_ i
89-5
*4
59*5
36-5
4 *o
—
__ 1
96*0
*5
49.0
41*0
10*0
—
~ |
90-0
16
42-5
36*0
20*5
1*0
__ 1
1
785
<7
6o-o
37‘5
2*0
o-s
97-5
37-16
8*64
079
0*02 1
l
90*52
451
The examination showed that there was a profound shift to the
left of the Arneth count in yellow fever. The index on the first days
represented in the collection of blood-films averaged 90* 5, and in
some cases actually 99 per cent, of the polymorphonuclear leucocytes
belonged to Classes I and II. Even after allowing liberally for a
possible personal factor in the counts, such a deflexion is much
greater than has been described by other observers in any other
disease. As in the severer cases of malaria, a great many of the
cells had a simple horseshoe-shaped nucleus, and in one patient
these forms constituted 82 per cent, of the polymorphs.
The shift to the left of the Arneth count in yellow fever is more
pronounced than it is in malaria. An attempt to show this fact
graphically has been made in Charts III and IV. Chart III shows
the average Arneth counts in yellow fever, in malaria, and in normal
persons in Europe. Chart IV shows the curves produced by plotting
the Arneth indices by percentages into ten equal groups in cases of
yellow fever, in Europeans suffering from malaria, and in
apparently healthy Europeans in Accra.
It has been shown above that in the twenty-three cases of malaria
studied at Accra in which the diagnosis was confirmed by the
finding of the parasites, the Arneth index averaged 86*6 as
compared with 90*5 in these seventeen cases of yellow fever. The
difference is one of degree; and as in some of the cases of yellow
fever the index was lower than it was in some of the cases of
malaria, the index alone could not be used as a means of making a
differential diagnosis between these two diseases from an examina¬
tion of the blood on the first day on which a case happened to be
seen.
In those cases of malaria, however, in which the evidence of
infection was presumptive only, the average index was found to be
79*3; and in the cases of fever merely suspected of being malarial,
it was lower still, namely, 71 ’35. It is in cases of the latter type
that it is important to be able to decide whether or not they are
yellow fever. The occurrence of a very high Arneth index in any
suspicious case of fever in which no malaria parasites could be
found, and in which there was not presumptive evidence of malaria,
would, I believe, be strongly in favour of a diagnosis of yellow
fever. It will be necessary, before arriving at a definite decision, to
452
study the condition in mild cases in natives, and to follow the
changes in the Ameth counts from the earliest stages of the disease
until convalescence is fully established. I hope to be able to do this
as soon as I can procure sufficient local material.
Chart III.—The average Arneth counts in (A) normal persons in
Europe (Arneth), (B) twenty-three cases of malarial fever at
Accra, and (Q seventeen cases of yellow fever
453
Chart IV.—An attempt to show graphically the degree of the shift to
the left in the Arneth counts in (A) healthy Europeans, (B) Euro¬
peans suffering from malaria, and (C) yellow fever.
454
IV. THE SIGNIFICANCE OF THE CHANGES OBSERVED IN THE
ARNETH COUNTS
A marked shift to the left of the Arneth count has generally
been assumed to be an indication of a lowered resistance. As the
result of a large number of examinations in various diseases, Arneth
concluded that there was a definite relationship between the blood
picture and the progress of the disease; and the view adopted by
him, and by most subsequent authors, was that the cells of Classes
I and II were younger, and less efficient than the cells of the higher
classes; and that therefore the degree of the shift to the left was an
index of the patient’s resistance. Chamberlain (1914) summarises
this view by saying ‘ a high Arneth index (excess of Classes I and
II) goes hand in hand with a low resistance, or with a high degree
of toxic and bacterial absorption which is bringing about the
destruction of the actively phagocytic leucocytes (Classes III and
IV).’
There is, however, little evidence in support of the assumption
that the neutrophile leucocytes of Classes I and II are less actively
phagocytic than those of the higher classes. Many observers have
failed to detect any difference in experiments carried out in vitro;
and I have recorded elsewhere a case of fatal malarial fever in which
the leucocytes had ingested a large number of parasites, making it
possible to gauge their phagocytic activity in vivo> and have shown
that in this case the polymorphonuclear leucocytes of Class I were
certainly not less efficient than those of the higher classes.
Breinl and Priestley (1914) have, therefore, advanced the view
that ‘ the Arneth picture is an expression of the functional activity
of the leucopoetic system, especially the bone-marrow, rather than
that of phagocytic activity.’ They dissent from the view of
Chamberlain and Vedder that the shift to the left observed in
Filipinos may be an indication of a lowered resistance to infections
on the part of the native races, and attribute the similar phenomenon
found in North Queensland children to purely climatic influences.
They state that in their experience there is amongst the children in
Townsville, who show a decided shift to the left of the Arneth count,
‘ no greater susceptibility to infectious diseases than amongst the
same class in Europe.’
+55
In West Africa the healing powers of the natives are famous,
and it is scarcely credible that the slight shift to the left of the
Ameth count observed in them can be an indication of a lowered
resistance to infection. As I have already stated, I believe that in
West Africa the shift in apparently healthy individuals may be
accounted for by infections with malaria parasites which do not
necessarily culminate in an attack of 'fever.*
The very pronounced shift to the left observed in confirmed cases
of malarial fever at Accra may be considered as supporting the view
of Breinl and Priestley that the phenomenon is an indication of the
functional activity of the leucopoetic system; for on this hypothesis
it is in such diseases as malaria, which profoundly affect the spleen
and the bone-marrow, that one might expect to find a marked shift
to the left indicating a morbid activity of these tissues. The Arneth
index was highest in those cases which showed the greatest
constitutional disturbances, and not necessarily in those that had
the largest number of parasites in the peripheral blood. In yellow
fever, another disease accompanied by severe constitutional
disturbances and a profound toxaemia, the Arneth counts showed a
shift to the left that was often extreme in degree.
The blood-films from yellow fever cases that were examined, were
some of those from which 0 *Brien (1914) described a degeneration
of the polymorphonuclear leucocytes which he considered to be
characteristic of the disease. Some of the abnormal forms he
interpreted, correctly I believe, as being dead cells. Identical forms
were described by Dr. Mary Rowley (1907) in a fatal case of aortic
and mitral disease with anaemia; and this author was able to bring
forward evidence in proof of the fact that these cells had been killed.
Similar polymorphonuclear leucocytes are seen occasionally in the
blood of normal persons; but they are relatively much more
abundant in acute cases of malaria and in yellow fever. It is
probable that in these diseases the profound toxaemia causes a great
destruction of the circulating polymorphonuclear leucocytes, and
thus leads to a relative increase in the peripheral blood of the young
cells—namely, those belonging to Ameth*s Classes I and II, and
a corresponding shift to the left of the count.
The increase in the percentage of young cells in the peripheral
blood might result from (1) the elimination of the cells of the higher
456
classes by the selective action of the toxins as suggested by
Chamberlain, or (2) a flooding of the blood with newly-formed
elements produced by the activity of the leucopoetic system. As a
leucocytosis is frequently observed in blood in which there is a
marked shift to the left of the Ameth count, and as there is really
no reason to suppose that the cells of the higher classes are the more
actively phagocytic, and so the more liable to be destroyed by a
high degree of toxic and bacterial absorption, I am inclined to think
that the latter explanation is the correct one.
It might be expected, then, that in an intense infection there would
be developed an immediate shift to the left of the Ameth count,
increasing rapidly in degree as the toxaemia deepened, and tending
to diminish as recovery took place.
SUMMARY
1. A slight shift to the left of the Arneth count is found in the
blood of healthy Europeans in West Africa.
2. There is a marked shift to the left in malarial fever which is
evident not only during the attack, but also before its onset, and
for a considerable time after convalescence is established.
3. It is probable that the abortive inoculations with malaria
parasites by infected mosquitos, which are a part of the daily life in
many parts of West Africa, are sufficient to account for this shift in
apparently healthy Europeans, without postulating a specific action
of the climate on the white races living in the tropics.
4. In yellow fever there is a great shift to the left of the Arneth
count.
5. It is suggested that the changes observed in the Ameth
counts are due to toxaemia causing a destruction of the circulating
polymorphonuclear leucocytes, and a flooding of the blood with
young cells liberated by the activity of the leucopoetic system.
REFERENCES
Buinl, A., and Priestley, H. (1914). Annals of Trop. Med. and Parasitol., Vol. VIII, p. 565.
Chamberlain, W. P. (1914). American Jour, of Trop. Diseases, Vol. II, p 48.
Cooke, W. E. (1915). Jour, of Path, and Bact., Vol. XIX, p. 494.
Knapp, H. H. G. (1915). Indian Medical Gazette, Vol. L, p. 95.
Macfie, J. W. Scott (1915). Lancet, May 1.
Rowley, Mary W. (1907). Jour, of Experim. Med., Vol. X. p. 78.
457
BABESIASIS AND TRYPANOSOMIASIS AT
ACCRA, GOLD COAST, WEST AFRICA
BY
J. W. SCOTT MACFIE, D.Sc., M.B.
WIST AFRICAN MEDICAL STAFF
(Received for publication 4 August , 1915)
Plates XXXV, XXXVI
CONTENTS
I. Babesiasis. page
A. Babesiasis of cattle and sheep . 457
B. Canine Babesiasis . 462
C. A piroplasm, NuttdUia decumani , n.sp., of the brown rat (Mus dtcumanus ) 462
II. Trypanosomiasis.
A. Trypanosomes found in blood films obtained from the Accra slaughter¬
house ... 464
B. Canine Trypanosomiasis . 477
C. Equine Trypanosomiasis . 478
D. A small monomorphic Trypanosome found in the blood of a mare ... 480
E. Two cases of Trypanosomiasis in mules resembling acute Do urine ... 486
BABESIASIS
A. BABESIASIS OF CATTLE AND SHEEP
In a previous paper (1914) I gave a preliminary account of the
occurrence of babesiasis in Nigeria, and described three species of
parasites found in the blood of cattle and sheep in that country.
Similar observations have been made by Bouet (1908) on the Ivory
Coast, and by Broden and Rodhain (1909) in the Congo; the former
recording the occurrence of P. parvum, P. mutans and P. bigeminum,
and the latter P. mutans and P. bigeminum. From these observa¬
tions it is evident that piroplasms are common on the West Coast,
and it may therefore be of some interest to record for comparison the
results of the examinations of domestic animals at Accra, in the Gold
Coast, since hitherto but little attention has been directed to these
parasites in British West Africa.
For the purposes of this investigation blood films were obtained
45 8
from the Accra slaughter-house. Hump-backed cattle, straight-
backed cattle, sheep, pigs, and goats are slaughtered daily at Accra.
Many of the animals are bred locally, but others come from a
distance, and no doubt bring with them their parasitic infections.
The hump-backed cattle, as in Nigeria, are bred in the north, and
are driven south in herds during the dry season until they reach the
coast towns. On the journey they are exposed to the attacks of ticks
and tsetse-flies, and debilitated by fatigue, they are probably but
feebly resistant to parasitic infections. The straight-backed cattle,
on the other hand, are mostly bred in the southern parts of the Gold
Coast, and the majority of those examined were either Accra or
Addah animals; they had not, at any rate, treked down from the
north like the hump-backed cattle. A considerable number of the
sheep had also come to Accra from a distance; but the
majority of the goats, and probably all the pigs, had been bred in
the neighbourhood.
A hundred animals of each kind were examined, namely hump¬
backed cattle, straight-backed cattle, sheep, pigs, and goats. Only
a single blood film was obtained from each animal, so that the
percentages of infections found are probably considerably below
those actually occurring. In this way piroplasms were found in
53 per cent, of the hump-backcd cattle, in 40 per cent, of the straight-
backed cattle, and in 21 per cent, of the sheep (Table I). None were
found in the pigs and goats.
Table I.—The results of the examination of 500 domestic animals for babesiasis.
Host
Number
examined
Number
, infected
with
Piroplasms
B. bigemina
T. mutans
Cattle, hump-backed breed .
100
1
53
7
49
Cattle, straight-backed breed .
100
40
3
38
Sheep .
100
21
—
21
Pigs.
100
—
—
—
Goats .
100
—
—
Totals.
500
114
10
108
459
So far as could be ascertained the infections appeared to be
benign, but as the animals had frequently concurrent infections with
other parasites, especially trypanosomes, it was difficult to determine
this point. For instance, forty-seven of the fifty-three hump-backed
cattle found to harbour piroplasms were infected with trypanosomes
also. In the case of the sheep no trypanosomes were found
associated with the piroplasms, and in these animals no clinical
symptoms of disease were observed.
Various abnormal conditions of the red corpuscles were found in
the blood films, notably a coarse basophilia. Anaplasma-like bodies
were present in many cases, but as these are common in healthy
animals they can be of no special significance in this connexion. In
the blood films from the cattle, especially the straight-backed breeds,
there were generally to be found oval or rounded bodies about the
size of a red blood corpuscle that stained a pale blue colour. A few
granules that stained similarly to chromatin were present in these
bodies. These basophile cells were similar to those described by
Castellani (1912) as occurring in man in cases of yaws, psoriasis,
lichen, acne, severe anaemia, &c.; and they were probably, as he
concludes, 4 red cells undergoing degenerative changes/ Although
piroplasms were present in the blood of most of the cattle in which
they were detected, there does not appear to be any direct connexion
between them and these parasites, since they were not constantly
associated. The basophile cells may, however, have been an
indication of the anaemia that is frequently induced by piro-
plasmosis, although, as has already been stated, the infections
appeared otherwise to be benign.
Two species of piroplasm were encountered in the blood films, the
one a large pyriform parasite, and the other a small oval or rod¬
shaped organism. Both types were found in cattle, and the latter
in sheep also. The third species found in Nigeria, a large parasite
with forms in which the chromatin was divided into six or seven
masses, was not found in the Gold Coast.
Owing to the lack of unquestionably uninfected animals for
experiments, it has not been possible to attempt to determine the
ticks that transmit these parasites. Experiments designed to decide
this point could not be conclusive if carried out with locally bred
animals in the Gold Coast because of the high percentage of natural
460
infections in the cattle and sheep, the benign nature of the infections,
and the extreme rarity of the parasites in some cases. Some ticks
collected from the animals were, however, sent home to England, and
were very kindly identified by Prof. Nuttall. The species from both
cattle and sheep were the same, namely, Boophilus y Ambylomma
variegatum , and Hyalomma aegyptium; and presumably one or
more of these transmits the piroplasms.
Babesia bigemina
The larger species of piroplasm appeared in the blood as oval or
pear-shaped bodies of considerable size. Pairs of pyriform bodies
were not uncommon, and when found were generally seen to occupy
the greater part of the enveloping erythrocyte. There can be little
doubt that these parasites should be identified as Babesia bovis or
bigemina (Piroplasma bigeminum). In the straight-backed cattle
the morphology of the parasites was similar to that found in dwarf
cattle in Nigeria (1914); but in the hump-backed cattle the pyriform
pairs were somewhat smaller, and their nuclear structure was less
distinct. In one hump-backed beast a considerable number of the
parasites were notably amoeboid.
This type of Babesia was found in 7 per cent, of the hump¬
backed cattle, and in 3 per cent, of the straight-backed breed.
They were never very numerous, and in four of the ten animals in
whose blood they were detected they were associated with the smaller
type of parasite to be described immediately.
Theileria mutans
The smaller type of piroplasm was identical with that found in
Nigeria. At Accra it was present in 49 per cent, of the hump-backed
cattle, in 38 per cent, of the straight-backed cattle, and in 21 per
cent, of the sheep. It was, therefore, a very common parasite of the
animals brought to the Accra slaughter-house, just as it was of the
domestic animals in Nigeria. In several of the hump-backed cattle,
and a few of the sheep, the infection was a heavy one; but in the
straight-backed cattle the parasites were as a rule scanty or rare.
Certainly in the majority of the animals the infection appeared to
be benign.
The parasites were very simple but remarkably pleomorphic
organisms, appearing as ring-shaped, oval, horseshoe-shaped, and
461
rod-like bodies, and characteristic cross-forms. Examples of all
these different forms were figured in my account of the Nigerian
infections, so that it will be unnecessary to illustrate the present
description. Since then I have had opportunities of examining
smears from the spleen in cases of several infected animals, but up
to the present I have not detected the presence of Koch's 'blue
bodies' in any of them. Such a parasite, according to the classifica¬
tion of Franca, would have to be included in the genus Theileria ,
and for this reason, in describing its occurrence in Nigeria, I
provisionally identified it as Theileria parva > stating at the same
time that Koch's 'blue bodies' had not been found, and that the
infections appeared to be benign. Minchin (1912), however,
considers that ‘ the diagnosis of the genus Theileria given by Franca
would appear to apply to B. mutans rather than to T. parva ,' and
states that 'a confusion has arisen between two parasites very
similar as regards the appearances they present in the blood, but
differing in every other respect, namely, Theileria parva , the true
parasite of " East Coast fever " of cattle, and Babesia (Piroplasma)
mutans , also found in cattle. In both parasites alike the charac¬
teristic cross-forms appear in the blood. In Theileria parva ,
however, the cross-forms are an aggregation of four distinct
gametocytes which have invaded the same corpuscle, while in
Babesia mutans the cross-forms are produced by quadruple fission
of an ordinary multiplicative individual.' The parasite described
from Nigeria, and that found recently in the Gold Coast, showed
cross-forms unquestionably produced by fission, and not by the
aggregation of four distinct gametocytes in a single cell. It should
therefore be identified, according to Minchin, as B. mutans. The
facts that the infections appear to be benign, and that Koch's ' blue
bodies' have not hitherto been detected in smears from the spleen
of infected animals, tend also to prove that the parasite is not
T. parva. Inoculation experiments would assist the diagnosis, as
T. parva is not inoculable; but for the reasons given above these
could not be conclusive if carried out in this country. At the same
time, an organism characterised by the occurrence of bacilliform or
rod-shaped parasites, and multiplicative forms in the shape of a
cross, cannot well be assigned to the genus Babesia , and it will
probably be best for the sake of clearness to identify this small
piroplasm as Theileria ( Babesia ) mutans.
462
B. CANINE BABESIASIS
Up to the present only a single case of canine babesiasis has
come under my notice at Accra. The dog found to be infected had
been brought into the Colony only six weeks previously. He was
feverish, much wasted, and very anaemic. The right eye was
dimmed, and there was a profuse watery discharge from the nose.
On examining the blood, large numbers of piroplasms were found;
many extremely amoeboid, but others showing the pyriform outline
and the association in pairs characteristic of Babesia cants.
Imported dogs are known to suffer on the Gold Coast from a
disease characterised by severe anaemia and rapid emaciation which
does not appear to be a trypanosomiasis. Possibly some of the cases
of this disease are due to babesiasis. It is the common experience
that in Accra dogs become infested with ticks, no matter how
carefully they are tended, and it would not therefore be surprising
if babesiasis were found to be a prevalent disease.
No case of equine babesiasis has as yet come under my notice
in the Gold Coast, although this disease is known to occur in
Nigeria and in the neighbouring colonies.
C. A PIROPLASM, NUTT ALU A DECUMANI, n. sp., OF THE BROWN
RAT (MUS DECUMANUS ). PI. XXXVI, figs. 1-14
In the blood of a few brown rats {Mus decumanus) that had
either been sent to me for examination by the Medical Officer of
Health for Accra, or had been caught in the laboratory itself,
unpigmented parasites were found in the erythrocytes. Four out
of twenty rats recently examined were found to be infected; that is,
a proportion equal to 20 per cent.
From two of the rats only a single blood-film was obtained, and
in both of these the parasites were rare. In the other two the
infections were heavier, and as I was able to keep the animals in
captivity, it was possible to make a more thorough study of the
morphology of the parasites. Both these rats were young animals,
and both showed in their red corpuscles, in addition to the piro¬
plasms, the bodies known as GrahameUa. The first was caught on
January 10th, 1915, and was kept under observation until February
15th, the thirty-seventh day, when it was accidentally killed. When
caught, the examination of the blood showed the presence of
4*3
Grahamella and Jolly bodies, but no piroplasms. A few days later
one or two piroplasms were detected, and by the fifteenth day a
considerable number were present, although the infection was never
a heavy one, and the parasites had always to be searched for most
carefully. From this date the parasites gradually diminished in
numbers, so that by the twenty-sixth day they were rare. They
were still present on the thirty-seventh day when the rat was killed.
The Grahamella bodies were not found after the thirteenth day.
The second rat was caught on April 27th, and is still under
observation. Piroplasms were found in the blood on the first day,
and continued to be present up to the tenth day, but have not been
detected since. They were always rare, and it was only after
prolonged search that they were found. The Grahamella bodies
have vanished from the blood of this rat also since it has been under
observation.
The forms of the parasite most commonly seen consisted of a
somewhat irregularly shaped chromatin mass, and an amoeboid or
ring-shaped protoplasmic body. The smallest parasite seen was a
minute dot of chromatin to which a very small ring of blue-stained
material appeared to be attached, but even at this early stage of
development there was a paler area or vacuole in the protoplasm
(PI. XXXVI, figs. 1 and 5). More mature parasites were often of
considerable size, and great irregularity of outline. The chromatin
was in many cases drawn out into a number of processes or separated
into two or more distinct masses, and the protoplasm was freely
vacuolated or extended in a number of delicate threads. Other
parasites were signet-ring-shaped, and closely resembled the
characteristic forms of Plasmodium falciparum . Sometimes there
were two chromatin dots at opposite sides of the ring. None of the
parasites was pigmented. The red corpuscles did not appear to be
affected by the presence of the parasites, and were neither obviously
enlarged nor stippled.
After long search a few specimens were found in which division
had taken place, resulting in the production of four lanceolate
parasites arranged in the form of a cross. Each of these bodies
contained a nucleus which was generally situated near the middle,
and in addition a second minute chromatin dot could usually be
seen at the distal end. No pairs of pyriform bodies were found.
Subcutaneous inoculations of blood containing these parasites
+64
were made into three white rats, but without transmitting the
infection. A single attempt to convey the parasites to another brown
rat, by means of ticks taken from the body of an infected animal,
was also unsuccessful.
This parasite would seem to belong to the genus Nuttallia , and
as I am unable to find any reference to a previous description of it,
I would suggest the name Nuttallia decumani for it, should it prove
to be a new species.
REFERENCES
Bourr, G. (1908). Piroplasmoie bovine observte A la c6te d’Ivoire. Bull . Soc. Path. Exot.,
Vol. I, p. 234.
Broden, A. and Rodhain, G. (1909). Piroplasmoses des bo vide* observees au Stanley-Pool.
Bull. Soc. Path. Exot., VoL II, p. 120.
Castellani, A. (1912). Note on Certain Cell Inclusions. Journ. Trop. Med. and Hyg., Vol.
XV, p. 354.
Macfie, J. W. Scott (1914). Notes on some Blood Parasites collected in Nigeria. Annals
of Trop. Med. and Parasit., Vol. VIII, p. 439.
Minchin, E. A. (1912). An Introduction to the Study of the Protozoa, p. 380. Edward
Arnold, London.
TRYPANOSOMIASIS
Three types of trypanosomes were met with at Accra, namely,
(1) a polymorphic parasite, (2) a large monomorphic parasite with
a long free flagellum, and (3) a short monomorphic parasite without
a free flagellum. These three types are the same as those found in
Nigeria, and which I have elsewhere identified as (1) T. fecaudi
(T. brucei of Uganda, T. ugandae ), (2) T. vivax , and (3) T. pecorum
( T. congoletise). In dried blood-films, such as those obtained from
the slaughter-house, morphological features had to be relied on for
the identification of the trypanosomes. Such data are, of course,
insufficient to distinguish the more closely allied species; but as
the small number of inoculations into animals that were made seemed
to confirm these diagnoses, they are probably correct.
A. TRYPANOSOMES FOUND IN BLOOD FILMS OBTAINED
FROM THE ACCRA SLAUGHTER-HOUSE
Blood-films were examined from 500 animals slaughtered at
Accra, namely, 100 each from hump-backed cattle, straight-backed
cattle, sheep, pigs, and goats. The films were the same as those
+65
used in the examinations for Babesiasis, of which an account has
already been given; and the remarks made there with regard to the
sources from whence the animals came should be taken into considera¬
tion with reference to the trypanosome infections. In Accra itself
tsetse flies are exceedingly rare. A few are caught every year, but
it is generally supposed that these have found their way into the
town in railway carriages or waggons. Within a few miles,
however, tsetse flies are numerous; and any animals coming from
a distance must be exposed to the attacks of these insects for the
greater part of their journey. It is probable, therefore, that the
majority of the trypanosome infections described had been con¬
tracted before reaching Accra.
Hump-backed Cattle. Trypanosomes were found in 92 per
cent, of the hump-backed cattle; and as only a single blood-film
was examined from each animal, this proportion is more probably
below the mark than above it. In the majority the parasites were
very numerous. Trypanosomes of the type of T . vivax were present
in 76 per cent., T . congolense in 28 per cent., and T. pecaudi in
12 per cent, (see Table II). Sixty-nine of the hundred animals
examined were infected with a single type of trypanosome, fifteen
with T. vivax and T. congolense , four with T. vivax and T. pecaudi ,
three with T. congolense and T. pecaudi , and one was the host of
all three types, T. vivax , T. congolense , and T. pecaudi .
Tablx II.—Trypanosdme infections found in animals killed at the Accra slaughter-house.
Number
Number 1 Percentages infected with
infected |
Host
examined
with i
trypanosomes T. vivax
T. congolense
T. pecaudi
Cattle, hump-backed
i
100
9 Z
76
j 28
! 1 2
Cattle, straight-backed
>00 !
18
14
6
1
Sheep .
100
4
3
2
-
Pigs.
100
5
—
5
—
Goats
100
1
1
1
Totals
500
120
• iX-8
8-4
2-6
466
Of all the animals slaughtered at Accra the hump-backed cattle
have the longest journey to come, and this fact probably accounts
for the great number of them found to harbour trypanosomes. Even
as far north as Kumasi, I understand, almost all are found to be
already infected with these parasites.
Straight-backed Cattle. Trypanosomes were found in 18 per
cent, of the straight-backed cattle. The parasites were generally
scanty, and were often detected only after a long search. T. vivax
was found in 14 per cent., T . congolense in 6 per cent., and
T . fecaudi in 1 per cent. Fifteen of the animals had a single
infection, and three harboured both T. vivax and T. congolense .
Straight-backed cattle are bred in the southern parts of the Gold
Coast, and many of those examined at Accra had not been brought
from any great distance. The relatively low percentage of trypano¬
some infections, and the rarity of the parasites in the blood, should,
probably, be correlated with this fact. Some of these animals were,
however, of the dwarf breed peculiar to West Africa, and it should
be remembered that, as I have suggested elsewhere (1913), they may
possesses a partial immunity to T . vivax at any rate.
SHEEP. Four of the hundred sheep examined were found to be
infected with trypanosomes, but in each case the parasites were rare.
T. vivax was found in 3 per cent., and T. congolense in 2 per cent.
Three sheep had a single infection, and one harboured both species
of trypanosome.
PlGS. Five out of the hundred pigs examined were found to be
infected with trypanosomes, and in each case the type of parasite
was T. congolense .* The parasites were generally very rare.
GOATS. Only one of the goats was found to harbour trypano¬
somes, and this animal had a double infection— T. vivax and
T. congolense. It is somewhat remarkable that so few infections
should have been found in goats, since these animals have been
credited with being a natural reservoir for T. vivax , the trypanosome
most frequently met with at Accra.
T. pecaudi ( T . brucei of Uganda, T. ugandae)
Polymorphic trypanosomes were found in thirteen animals
(2 6 per cent.), namely, in twelve hump-backed cattle, and in one
• Subsequently a pig was examined in who?e blood trypanosomes of the T. pecaudi type were
present.
4 6 7
straight-backed cow. This type of trypanosome was the most
uncommon of the three forms occurring at Accra. It is the most
fatal to domestic animals in Nigeria, and so probably in the Gold
Coast also. In the hump-backed cattle it was often associated with
other trypanosomes: four times with T. vtvax , four times with
T. congolense , and once with both T. vtvax and T. congolense.
Morphologically the trypanosomes resembled T. pecaudi or
T. brucet of Uganda, both long and slender, stumpy, and inter¬
mediate forms were found in each case, but forms with posterior
nuclei were not seen. It is impossible, therefore, to exclude the
possibility that some of the trypanosomes might have been
T, gambiense. The occurrence of posterior nuclear forms is,
however, very irregular in cattle infected with trypanosomes which,
when subinoculated into rats and guinea-pigs, show high percentages
of these forms; and at the periods during which the parasites are
scanty, as they were in all the slaughter-house cases, it is generally
impossible to find them at all. Too much importance should not,
therefore, be attached to the absence of these forms, especially when
the materials examined are restricted to dried blood-films.
In its behaviour in the tsetse fly, T. pecaudi is said to differ
from the other polymorphic trypanosomes, development taking place
in the gut and proboscis, instead of in the gut and the salivary
glands. Confirmation of this observation is desirable; but in any
case, in dealing with these slaughter-house films the exogenous cycle
of development had to be ignored, and it was impossible to carry
identification beyond the point at which the trypanosomes were
assigned to what is sometimes known as the ‘ T . brucei group.*
In one instance, blood containing this type of trypanosome was
obtained from a hump-backed ox, and inoculated subcutaneously
into a white rat and a guinea-pig. The rat first showed trypano¬
somes in its blood on the tenth day, and death took place on the
nineteenth day. The parasites were at first very scanty, but
gradually increased in numbers up to the day of death; and in the
latter stages of the disease, forms with posteriorly placed nuclei were
numerous. The guinea-pig did not become infected, but with two
others was successfully inoculated with blood taken from the rat.
In these three guinea-pigs the incubation periods were respectively
twenty, nineteen, and seven days; and the duration of the disease
thirty-four, twenty-five, and thirteen days.
468
Measurements were made of 200 trypanosomes from the blood of
the rat (see Table III, and Chart I). The average length was
27 66 p; the longest individual measuring 39 p, and the shortest
15 M- This trypanosome then corresponded with T. pecaudi
(T. brucei of Uganda), both in its morphology and in its patho¬
genicity to rats and guinea-pigs.
Table III. —Measurements in length of T. pecaudi (cattle strain).
Animai
Day of
the
disease
Number
measured
Length in microns
Average Minimum I Maximum
!
Rat .
1
2°
Rat
2
2 5
Rat .
3
20
Rat .
4
2 5
Rat .
5
. 20
Rat .
6 1
2 5
1
Rat .
7
20
Rat .
8
2 5
Rat .
9
20
'i
200
*7-65
16
33
2 77 2
! «7
37
26*25
16
35
3 2 *64
26
|
39
26*65
1
18
33
26*88
16
35
28*2
16
38
27-2
‘5
39
24-8
16
35
27*66
*5
39
4<*9
470
T. vivax
Trypanosomes morphologically identical with T. vivax were
present in ninety-four (18*8 per cent.) of the animals examined,
namely, in seventy-six hump-backed cattle, fourteen straight-backed
cattle, three sheep, and one goat. They caused by far the greatest
number of the trypanosome infections met with at Accra, and in
many of the animals, especially the hump-backed cattle, were
present in the blood in enormous numbers.
Blood from hump-backed cattle heavily infected with these
trypanosomes was inoculated into two white rats, two guinea-pigs,
and one rabbit, but without result. The insusceptibility of the
smaller laboratory animals, taken in conjunction with the very
characteristic morphology of the parasites, is sufficient evidence for
the identification. Measurements were, however, made of one
hundred trypanosomes, fifty from hump-backed cattle, and the same
number from straight-backed cattle (see Table IV). The average
length was found to be 22 64 /x, and the range from 18// to 29 yu.
These measurements, and the curve shown in Chart II, are
consistent with the identification T. vivax.
Table IV.—Measurements in length of T. vivax from cattle.
Host
Number
measured
Distribution according to length in microns
Average
18
O
21
22
23
24
26 ; 27
28
29
Cattle, hump-backed, No. 451 ...
25
I 2
+
4
8
5
1
!
—
—
22*40
Cattle, hump-backed, No. 630 ...
*5
I 1 —
I
4
9
6
1
2 1
—
—
23*32
Cattle, straight-backed, No. 589
25
1
— 1 2
2
5
5
3
4
1 1 1
—
1
23*16
Cattle, straight-backed, No. 734
25
—
— I
5
5
7
4
3
~~ “
—
—
22*68
100
1
'
2 5
1
12
18
29
1-
9
3 2
—
1
22*64
47i
Chart II.—The distribution, by percentages, in length of T. vivax
from cattle.
472
T. congolense
Small monomorphic trypanosomes with a scanty undulating
membrane, and without a free portion to the flagellum, were present
in forty-two (8 4 per cent.) of the animals examined, namely, in
twenty-eight hump-backed cattle, six straight-backed cattle, two
sheep, five pigs, and one goat. The infections were never very
heavy, and in several of the hosts only one or two individuals were
found in a large blood-film.
Both a guinea-pig and a white rat were on two occasions
inoculated with blood containing these small trypanosomes, but
with negative results. It was unfortunately impossible to experiment
with larger animals, such as goats, sheep, and cattle, and no dogs
were available. No detailed measurements at known periods of the
infection could therefore be made. Fifty individuals, taken as they
came, were, however, measured in blood-films from hump-backed
cattle and pigs naturally infected with these parasites (see
Table V). The fifty from cattle averaged 12*941* in length; the
longest measuring 17/* and the shortest 10 a*. The average breadth
was 2*29 a*, the broadest being 3*5/1 and the narrowest 1*25 a*. In
pigs the average length of the fifty trypanosomes measured was
12 24A*, the longest being 16/1 and the shortest ioa*. In breadth
they averaged 199/*, and ranged from 3*25/* to 125/*. The
distribution according to length and breadth of the hundred
individuals is shown in Charts III and IV.
The identity of these trypanosomes is open to doubt. Similar
specimens from horses which were taken to England from Nigeria in
1911 were identified by Bruce as T. pecorum , a species which is
apparently indistinguishable from T. nanunt , and which, according
to Blacklock and Yorke (1913), is probably identical with
T. congolense. The trypanosome found in the Gold Coast is
morphologically similar to that found in Nigeria, although in both
colonies the parasite was found to vary from the type to a consider¬
able extent. In experiments conducted at Eket this variation was
found to occur in the same strain when transmitted to different
hosts. There seems to be some reason to suppose that T . fecorum>
T . nanunt , and T. congolense are all strains of the same species of
trypanosome; in which case the name T. congolense is that properly
applicable to it.
473
With regard to T. dimorphon , such uncertainty exists as to the
nature of the original strain that it is doubtful if the name should
now be employed. The original figures given by Dutton and Todd
(1903) of their Gambian Horse Trypanosome show two types of
parasite that would certainly to-day be identified in Nigeria or the
Gold Coast as T. congolense (T. nanuni ) and T. pecaudi (T . brucei
of Uganda). An infected horse sent home some years later by
Table V.—The measurements in length and breadth of T. congolense ( T . nanum ).
Length in microns Distribution according to length in microns
Host Number _ _
measured
Max.
Average
1 Min.
i
8
9
10
11
12
>3
14
*5
16
17
18
Pigs.
... 50
16
12-24
| 10
—
—
2
11
*3
9
7
6
2
—
—
Cattle, humped
50
*7
I2 ’94
IO
—
—
9
2
10
9
11
3
3
3
—
100
l 7
! 2-59
10
—
—
11
1 13
2 3
18
18
9
5
3
—
Host
, Number
Breadth in Microns
Distribution
according to breadth in microns
measured
Max.
1 Average
Min. 1
1 * ^
«-5
i *75
2
2-25
1.5
2-75
3
3* 2 5
3*5
Pigs.
... 50
3*25
i -99
1-25
—
4
4
" 1
*4
8
3
4
—
1
—
Cattle, humped
... 50
3-5
2*29
1*25
—
1
3
3
! 5
5
12
2
6
2 1
1
100
3-5
|
2-24
1 *25
5
7 '
15
2 9
*3
*5
6
6
_j
3
1
Dr. Todd as probably harbouring T. dimorphon proved on investi¬
gation by Yorke and Blacklock to have a double infection with
T. congolense and T. vivax. It is probable, therefore, that the
animals originally examined by Dutton and Todd were infected
with all the three types of trypanosome common in West Africa,
namely, those referred to here as T. vivax , T. congolense , and
T. pecaudi. In the sense in which the name is used by Laveran and
Mesnil, T. dimorphon denotes a polymorphic species of trypano-
474
Chart III.—The distribution, by percentages, in length of T. congolense
(T. nanutri).
475
Chart IV.—The distribution, by percentages, in breadth of T. congoUnse
(T. nanum ).
476
some, highly pathogenic to the smaller laboratory animals, all the
individuals of which are without a free flagellum, and which shows
forms measuring from 11 fi up to 30 fi in length. Such a trypanosome
I have not yet seen in West Africa. Mesnil (1915) considers that the
small monomorphic strains of trypanosome isolated by me at Eket
from G . tachinoides were T. dtmorphon , apparently because a single
individual was found to attain the length of 21//. Two strains were
isolated by me, and in the second a single trypanosome was
measured that reached this length. This strain I noted in my
description was unusually large, but I did not separate it off from
the other and more typical one because I observed considerable
variations in the size of the latter parasite when inoculated into
different animals. The trypanosomes of the first and more typical
strain measured from 9/1 to 18/*, and averaged 13*9/* in length;
and I think it could not but have been T. congoletise. If the second
strain were not a variety of the same parasite, as I believe it to have
been, it must, I think, have been a new species. Its length, average
15’ 15A 1 , maximum 21/1, minimum n /*, does not approach that of
T. dimorphon.
The ideal relationship between a parasite and its host would
appear to be the benign infection, since a strain that kills its host
commits racial suicide. In the case of these trypanosomes an
approximation to the ideal relationship exists in game, and in
certain domestic animals. It might be expected, however, that a
parasite specialised so as to produce a benign infection in a certain
type of host might show wide variations in pathogenicity when
introduced into other animals. The smaller laboratory animals,
such as rats, guinea-pigs, and rabbits, in which the pathogenicity
of trypanosoifies is generally studied, are species that tsetse flies
normally would seldom have an opportunity of infecting, and it
might therefore be expected that irregular variations would occur in
them as the result of artificial inoculations. The significance of the
fact that T. congoletise is said to differ from T . nanum only in the
fact that it is pathogenic to these smaller laboratory animals, is
therefore of doubtful value.
Strains of trypanosomes that have been carried on for a number
of years by inoculations from animal to animal in a laboratory, in
unnatural surroundings, often in unnatural hosts, and without the
natural alternation of development in the insect (exogenous cycle)
477
and in the vertebrate (endogenous cycle), might be expected to show
. abnormalities. One direction in which abnormality might be
anticipated would be in the pathogenicity, and it would be most
probable that this would take the form of a, loss of specificity. This
has actually been found to be the case by some observers.
Blacklock and Yorke (1913a), for instance, succeeded after a
number of passages in infecting rabbits with T. vivax, a species of
trypanosome that is usually innocuous to these animals. The same
observers found that the early inoculations into laboratory animals
of a small trypanosome isolated from a horse infected in the Gambia
mostly failed, whereas the later were invariably successful.
If such alterations in pathogenicity take place in laboratory
strains, it is probable that they also occur occasionally in nature.
This might be the result of a natural variation in the virulence of
the trypanosome, of some idiosyncrasy of the host, or of both these
factors. Some such explanation has indeed been advanced to
account for the human infections with polymorphic trypanosomes
characterised by posterior nuclear forms (T. rhodesiense) which occur
in some countries, but apparently not in others in which parasites
morphologically identical are common in game and domestic
animals.
For the above reasons it seems probable that differences in
pathogenicity to the smaller laboratory animals are not of specific
importance, although in some localities they may be so constant as
to constitute distinct ‘ races.' And as regards T. congolense
( T . pec or uni) and T. nanunt, that are morphologically identical, that
infect the tsetse fly in the same manner, and that differ only in their
pathogenicity to laboratory animals, I agree with Blacklock and
Yorke (1913b) that in the present state of our knowledge we can
only conclude that they are the same parasite.
B. CANINE TRYPANOSOMIASIS
Only one dog was examined in whose blood trypanosomes could
be found. This animal, a bull-terrier, showed a very small
infection with small parasites of the T. congolense type. The
trypanosomes were too rare for it to be possible to study their
morphology by means of measurements; but the few individuals that
478
were found were similar in shape and size to the trypanosomes of
this type found in cattle and horses. The dog was in a very poor
condition, much wasted, covered with sores, and with a profuse
watery discharge from the eyes. The disease ended fatally.
C. EQUINE TRYPANOSOMIASIS
Nineteen cases of equine trypanosomiasis were met with in the
nine months during which the present investigation was in progress,
namely, in thirteen horses, five mules, and one donkey. Seven of
the horses were infected with T. pecaudi ( T . brucei of Uganda),
four with T. congolense ( T . pec or urn), and two with T . vivax (see
Table VI). All the five mules were infected with T. pecaudi
(T. brucei of Uganda), and the donkey with T. congolense
(T . pecorum).
Table VI.—Equine trypanosomiasis at Accra.
Cases of
Number infected with
Host
1 somiasis
seen
T. vivax
T. congolense
T. pecaudi
Horses .
13
2
! 4
7
Mules .
5
—
—
i 5
Donkeys ... ... ... ...
1
i
—
i
—
!
Totals . j
19
i
1
2 1
! '
5 !
12
All the animals belonged to Accra, and had not been out of the
town for periods varying from two or three months to over ten years.
It is practically certain, therefore, that they had been infected
actually in Accra itself, either by stray tsetse flies, or by some other
biting insect such as a Stomoxys or Lyperosia .
Mules, which are generally imported from the Canary Islands,
479
are largely employed in Accra, as they are thought to be less
susceptible than horses to trypanosomiasis. The immunity, if it
exists, can only be a very partial one, and does not protect them
from fatal infections with T. fecaudi.
The trypanosomes found in horses in Accra were the same as
those occurring in cattle, and the remarks already made with regard
to the identification of the latter apply also to the former.
One of the horses, however, was infected with a small trypano¬
some of the T. congolense type that differed in some respects from
the other parasites of this group found in domestic animals at Accra.
A short description of this parasite, and of the clinical features of
the disease caused by it, is given below.
T. pecaudi (T. brucei of Uganda) is almost invariably fatal to
horses, T. vtvax almost never so, whilst T. congolense ( T . nanuni)
has a virulence intermediate between these two. It must be
remembered, however, that even T . vivax is a serious scourge, since
it incapacitates its victims for a very long time.
Trypanosomiasis in Accra should be an easily preventable
disease. Tsetse flies do not appear to breed in the immediate
vicinity of the town, and the specimens occasionally captured within
its bounds are supposed to have been brought in on trains, waggons,
motor cars, etc. Occasional cases of trypanosomiasis might therefore
be expected, but there would appear to be no reason whatever why
there should be so many annually as occur at present. No
precautions are taken to prevent the spread of the disease, and
animals heavily infected with trypanosomes are allowed to move
freely about the town. As, however, it is well known that trypano¬
somes can be transmitted, at any rate mechanically, by Stomoxys ,
and other blood-sucking insects besides Glossina , it would perhaps
be vain to expect any reduction in the number of cases in horses and
mules in Accra so long as herds of cattle are allowed to loiter about
the streets, and to graze wherever they can find a patch of grass.
The cattle carry with them wherever they go various biting flies,
and as has been shown above, 92 per cent, of the hump-backed
cattle and 18 per cent, of the straight-backed breeds are infected
with trypanosomes of the same species as those that infect horses
and mules.
480
D. A SMALL MONOMORPHIC TRYPANOSOME FOUND IN THE
BLOOD OF A MARE. PI. XXXVI, figs. 15-38
One strain of trypanosome of the T. congolense group, found in
a horse, presented certain morphological features which were
unusual, and a short description of this parasite may not be out of
place.
The animal in which the infection was found was a mare brought
to Accra from England some years ago. She had not been out of
Accra for at least eight months before the commencement of her
illness, so that it is probable that the infection was contracted in
Accra. On the 24th January, 1915, she was sent to the laboratory
for examination, because one day she had stumbled and nearly
fallen down when being driven, and because it was noticed that her
joints were swollen. The first indications of illness had been
observed two or three days before this date.
On examination, the mare was found to be in good condition,
but there was some oedema of the legs, and in the blood a few
trypanosomes were found. During the next few days the trypano¬
somes increased in number; but on the 5th February they were
again scarce. On the 6th February, two localised patches of
oedema, each about the size of the palm of the hand, appeared on
the left side; the one in the saddle area, and the other a little
further back. Three days later these raised patches or plaques had
almost entirely vanished.
Trypanosomes continued to be constantly present in the blood,
but were never plentiful; and the mare gradually lost condition
until, by the end of February, she was obviously extremely ill. On
the 4th March she became suddenly much worse: the oedema of
the legs became considerable, the anaemia very marked, the hind
feet began to drag badly, some discharge from the eyes appeared,
and all over the body there appeared a large number of little raised
patches of oedema, each about the size of a shilling or a little larger.
These patches or plaques could be distinctly felt as raised discs with
a more or less circular outline, and a definite edge. The hair over
them was slightly roughened. The trypanosomes were also more
numerous in the blood, but they were still not abundant.
Up to this time treatment with injections of Atoxyl, and
4 8
arsenious acid and perchloride of mercury by the mouth had been
employed; but as these drugs had not produced any perceptible
benefit, on the 8th March, at the request of the owner, a native
horse-doctor was given the chance of proving the efficacy of his
methods. On the 21st March, however, it was reported that the
‘ cure 1 had failed, and that the mare was much worse. I saw her
the following day, and examined her blood, and found trypano¬
somes still present, but not numerous. She was then obviously
dying, and was unable to leave her stable. She was greatly wasted,
her skin was covered with small patches of oedema or plaques, the
oedema of the legs and abdomen had increased greatly, there was
profuse diarrhoea, and discharges were running from the eyes, nose,
and vagina. On the following afternoon, March 23rd, the mare
died, the disease having lasted just two months.
The Morphology of the Parasite
The living, unstained trypanosomes were short, and not notably
active organisms. They did not show any marked tendency to
progress, but, like so many of the trypanosomes belonging to this
group, were often seen buried between clumps of red blood corpuscles.
When fixed and stained the parasites were seen to be short,
rather stumpy, monomorphic trypanosomes. They were studied in
films stained by Leishman’s method, and their general appearance
is shown in Plate XXXVI. The ratio of breadth to length was
usually 1 to 6*8. The posterior end was pointed, or sub-acute. The
anterior end tapered rapidly in front of the macronucleus, somewhat
in the manner characteristic of T. vtvax. The micronucleus was
small, occasionally terminal, but more usually lying some distance
from the pointed posterior end, and sometimes situated laterally.
The macronucleus was oval, and was frequently situated at the
extreme anterior end of the body (PI. XXXVI, figs. 18, 21, and
22). On some days this remarkable position of the macronucleus
was observed in 50 per cent, of the trypanosomes. The undulating
membrane was poorly developed, and was, indeed, seldom visible
at all. The flagellum was without any free portion. The cell
contents were sometimes homogeneous or showing a finely reticulated
structure, and sometimes contained a few granules posterior to the
macronucleus. In the final stages of the disease, many of the
482
trypanosomes showed a single deeply stained chromatin dot
immediately posterior to the macronucleus. This body closely
resembled a second, and larger, micronucleus in its appearance and
in its reactions to stains, but the parasites in which it was seen were
not dividing forms. This feature was not peculiar to this strain.
Similar bodies have been seen in other trypanosome infections. But
as they were a most conspicuous feature in this case, and as they
appear in the figures, they should be mentioned.
Measurements were made of the length of three hundred trypano¬
somes (see Table VII and Chart V), fifty being taken as they came
Table VII. —Measurements in length of a small monomorphic trypanosome found in a mare.
Date
Number
counted
Length in microns
Distribution according to length in microns
Max.
Average
Min.
8
9
10
11
12
*3
*4
15
16 17
January 24 .
50
16
1278
10
—
—
3
7
h
10
7
8
(
1 1 —
February 23 .
50
15
I2*IO
9
—
.
5
8
18
12
4
2
— —
February 26 .
50
15
IP98 1
8
•
2
4
12
12
11
6
2
March 4 .
5 °
16
12*98
8
1
1
3
3
8
14
11
7
2 | —
March 8 .
5°
16
12*66 ;
9
_
■
5
1 3
*3
15
6
6
1 1 —
March 22 .
5 °
17
12*64
8
1
—
8
5
9
9
9
7
* 1 1
300
17
12*52 j
8
3
1 _
5
L
|
0*
00
74
71
! 43
32
5 j *
4 8 3
Chart V.—Distribution, by percentages, in length of a small
monomorphic trypanosome found in a mare.
4 8 +
on each of the six days on which it was possible to find this number
in the blood-films. The average length was 12*52//, the longest
individual measuring 17//, and the shortest 8//. The breadth at the
widest point was measured in 275 of the same trypanosomes. The
average was i'84/z, the broadest measuring 3*5//, and the narrowest
ro n (see Table VIII, and Chart VI).
Table VIII.—Measurements in breadth of a small monomorphic trypanosome found in a mare.
Date
January 24 ...
February 23
February 26
March 4
March 8
March 22 ...
Number
counted
Breadth in microns
Distribution according to breadth in microns
Max.
Average
Min.
1
1 " 2 5
*‘5
175
2
2 * 2 5
2 ‘5
2 *75
3
3* 2 5
3*5
5 °
3 -o
1-83
,•0
2
2
*3
9
6
3
—
1
—
-
2 5
3-0
2>I 5
i * 2 5
—
1
2
3
5
4
3
3
4
—
-
50
3 -o
i*8o
1*0
2
4
11
9
,8
4
—
1
1
—
-
5 o
3*5
1-85
p2 5
7
9
6
21
4
1
1
—
—
1
5 °
2 ‘75
1-87
I<2 5
—
4
9
9
19
5
3
1
—
—
-
50
3 -o
i *73
1 *0
6
6
12
6
3
4
1
1
—
—
2 75
3*5
1-84
1*0
10
24
56
42
, 88
I
_ 26 i
.4
7
7
— 1
1
The susceptibility of experimental animals
Owing to the scarcity of animals, inoculations with this trypano¬
some could only be made into three white rats, two guinea-pigs, and
one rabbit. None of these animals became infected.
Identification
The morphology and the measurements of this trypanosome
clearly show its affinity to the species T. congolense sensu lato;
that is, the T . pecorum of Bruce, the synonyms for which, he says,
are T . confusum (Kinghorn and Montgomery) and T . nanum
(Laveran), and to which T. dimorphon (Laveran and Mesnil) and
T . congolense (Broden) are closely allied species. In consequence
of the failure to infect rats, guinea-pigs, and a rabbit in the few
attempts made, it would probably be considered as most nearly
48s
Chart VI.—Distribution, by percentages, in breadth of a small
monomorphic trypanosome found in a mare.
486
allied to T . nanum by those who adhere to the view that this species
is valid.
The morphology of the parasite, however, showed certain
peculiarities. The average length, 12 52/i, is considerably less than
that given for either T. nanum or T. pecorum, although the trypano¬
somes were studied in thin blood-films that had been allowed to
dry, a procedure which is said to tend to increase their dimensions.
Bruce (1914) gives the average length of T . pecorum as 14// , and in
an earlier paper (1911) as 13 3 fi; and Kinghorn and Yorke (1913)
give the figure 13*6 n. The average length of the monomorphic
trypanosomes of this type which I isolated from G. tachinoides at
Eket in Nigeria (1914) was also considerably greater, namely 141/x.
The range of length, 8yu to 17/i, is, however, very much the same
as that given by most authors for trypanosomes of this species
( T . congolense in its wide sense); and this is probably of greater
importance than the average, since the latter may be subject to
variations from a number of causes, and certainly shows marked
differences, even in the same strain, when inoculated into different
animals.
The peculiar position of the macronucleus, at the extreme
anterior end of the parasite, was a most conspicuous characteristic
of the trypanosome, and could not, I think, have been due to the
technique employed in making the blood-films. I have not been
able to find any reference to this peculiarity in the descriptions of
any other similar trypanosomes. The occurrence of these forms
would seem to mark off this trypanosome as at any rate a well-
defined variety. The clinical aspect of the disease in the original
host was also unusual. The appearance of raised disc-like patches
or plaques on the skin was peculiar in my experience in West
Africa. For convenience in reference I suggest, therefore, that this
trypanosome should be named Trypanosoma congolense , var.
equinutn.
E. TWO CASES OF TRYPANOSOMIASIS IN MULES RESEMBLING
ACUTE DOURINE
Quite recently, two mules have come under my notice suffering
from an infection with a polymorphic trypanosome of the type
T. pecaudi (T. brucei of Uganda). In both these animals the
+®7
remarkable features were the rapidity with which the disease
developed, and the appearance shortly before death of small patches
of oedema on the skin.
The first of the two mules (see PL XXXV, fig. i) was brought
to me on the afternoon of June 16th. It was hardly able to move,
the hind legs being stiff and paresed; the head had to be supported
or the animal would have fallen, the eyes were closed, and there was
a watery discharge from the eyes and the nostrils. Oedema was
not conspicuous, but the legs were a little puffy. Trypanosomes
were found in the blood, but they were very scanty. The animal
was somewhat wasted, and looked as if it were in the very last
stages of trypanosomiasis; and it was hardly credible that it could
have been at work and apparently in its normal health the same
morning, which nevertheless was the case. The next day the mule
was even worse, and stood in the stable with its head bent almost
to the ground, and resting against the wall, its hind quarters
drooping, and the right hind hoof turned backwards from the
fetlock. On this day, all over the skin of the neck and sides a
number of small plaques were observed, each about the size of a
shilling; and the general oedema began to increase especially in the
dependent parts of the face. On June 19th, the third day after the
onset of the symptoms, the animal was in such a distressing
condition that it had to be destroyed.
The second mule was first seen on June 17th. In the morning
it had been apparently quite well, and had done its usual round of
work. At 3 p.m., when I saw it, it was in very much the same
condition as that described above in the case of the first mule. The
gait in particular was remarkable, and suggested irresistably that
the animal was extremely drunk. The plaques on the skin (see
PI. XXXV, fig. 2) were first seen on June 18th, and on the 19th the
animal fell down, and had to be killed, as it was unable to rise again.
In neither of these animals were the trypanosomes numerous in
the blood, but sufficient were examined to prove that they were
morphologically identical with the type of parasite found in cattle
and horses in Nigeria and the Gold Coast, and which is referred to
in this paper as T . fecaudi.
Owing to the lack of experimental animals, it was only possible
to inoculate a single guinea-pig from each of the infected mules.
488
The guinea-pig inoculated from the first mule did not become
infected. That inoculated from the second mule first showed
trypanosomes in the blood on the seventh day, and died on the
ninth day. Another guinea-pig inoculated from it died two days
later from some unknown cause, and as the mule itself had died in
the meantime, the strain was unfortunately lost. In the blood of the
single guinea-pig that became infected, the trypanosomes were
numerous on the last two days, and forms with posteriorly placed
nuclei were common. The measurements in length of this trypano¬
some in the one mule in which it was possible to find twenty-five
parasites in the slides taken, and in the guinea-pig infected from it,
are shown in Tables IX and X. The average length was 22*39 fi y
the longest being 33 fi, and the shortest 13 jjl.
The rapidity with which the symptoms developed in these two
cases was unique in my experience. The course of the disease
caused by the polymorphic trypanosome commonly found in Nigeria
and the Gold Coast (T. fecaudi ), although usually rapid, has
always been a matter of weeks or months in the horses and donkeys
that I have examined. The onset has always been rather gradual,
the first indications of illness being a loss of vitality, and a tendency
to stumble, and the symptoms then slowly developing, some cases
terminating fatally in two to four weeks, others lingering on for as
many months. Laveran and Mesnil (1912) state that the duration
Table IX.—Measurement# of 75 trypanosomes of the polymorphic strain found in
two mules at Accra.
1
1
Length in microns
Host
Number
measured
Day of the infection
Average
Mini¬
mum
Maxi¬
mum
Mule No. 923
2 5
Natural infection. Two days
before death
1
20-76
13
3 2
Guinea-pig No. 79 ...
2 5
Inoculated from Mule No. 923.
First day of infection
25-14
l 7
3 2
Guinea-pig No. 79 ...
2 5
Inoculated from Mule No. 923.
Second day of infection, and
day before death
21-28
16
33
1
22-39
13
33
489
Table X. —Distribution according to length of
two mules at Accra.
Number
Host 'measured Day °* infection
*3 *4
Mule No. 923 ... 25 Natural infection. Two 1 1
days before death
Guinea-pig No. 79 ... 25 Inoculated from Mule — —
No. 923. First day
of infection
Guinea-pig No. 79 ... 25 Inoculated from Mule ^ — —
No. 923 Second day
of infection, and day
before death
Totals . 11
; Number , . '_
Host measured Day of the infection
24
Mule No. 923 . 2$ Natural infection. Two 3
days before death
Guinea-pig No. 79 ... 25 Inoculated from Mule 2
No. 923. First day
of infection
Guinea-pig No. 79 ... 25 Inoculated from Mule —
No. 923. Second day
of infection, and day
before death
Totals
5
of the polymorphic strain found in
Length in microns
15 16
OO
19 20
N
N
1
1
—
2 —
* 3
I 2
I I
1 |
3 4
4 3
2 2
3 3
4 *
7 6
Length in microns
49 °
of the disease due to T. pecaudi is three to four months. In no
other cases have I seen such a dramatic onset as occurred in these
two mules, which were apparently well in the morning, and obviously
dying the same afternoon.
Neither have I previously seen the little patches of oedema of
the skin in infections with polymorphic trypanosomes in West
Africa. These patches were raised, and rounded, and about the
size of a shilling or a little larger. Over them the hair bristled
(see PI. XXXV, fig. 2). They answered very closely to the
description given by Pease of the plaques in dourine, as looking as
though a metal disc had been slipped under the skin. Such plaques
are generally considered to be characteristic of dourine (T. equiper-
duni) y and indeed the general appearance of the animals, and their
attitude, which almost exactly resembled that shown by Laveran
and Mesnil (1912) in their fig. LXXIII, suggested this disease.
Cazalbou, however, has described similar cutaneous lesions in
infections with T. pecaudi , but Bouffard did not observe them, and
Pecaud considered that they were rare. In another part of this
paper I have described the case of a mare infected with trypano¬
somes of the T. congolense type, in which similar dermal plaques
were a prominent symptom. Plaques are not therefore pathognomonic
of dourine, although they must still remain an important sign for
the purposes of differential diagnosis.
Dourine does occur in an acute form, as described by Laveran
and Mesnil, in which ‘ a Tengorgement du d6but, succedent une
paralysie aigue soudaine ou des acc&s de vertige qui emportent le
malade en quelques jours *; and this account corresponds with that
given above of the course of the disease in these two mules. The
morphology of the trypanosome found in these cases was moreover
not incompatible with a diagnosis of dourine, since of the four strains
of T . equiperdum examined by Blacklock and Yorke (1913) one, for
which they proposed the name T. equi , was found to be dimorphic,
and indistinguishable from T . rhodestense.
After most careful enquiries, however, it was proved conclusively
that neither of the mules could possibly have contracted the disease
in coitus. Both were mares, and had been in Accra for five and
three years respectively, during which time they had not been
covered. Unless, therefore, T. equiperdum {T . equi) can also be
conveyed by some other means, such as the bites of insects, which is
+ 9 1
not altogether impossible, these mules cannot have been infected with
this species of trypanosome.
So far as I am aware, dourine (Mai du Coit) is not known to
occur in the Gold Coast. I am convinced, nevertheless, that the
infection in these two mules cannot have been the same as that due
to the polymorphic trypanosome of which I had the opportunity of
studying a large number of cases in horses in Nigeria. I am
inclined to think that the trypanosome may have been the same as
the Runcorn Laboratory strain of T. equiperdum described by
Blacklock and Yorke (1913), which was one that had been brought
from Algiers in a horse by the firm of Hagenbeck. This was the
dimorphic strain indistinguishable from T. rhodesiense , for which
the name T. equi was proposed. In the present uncertainty as to
the identity of mammalian trypanosomes, it is impossible to
determine what is the connexion between this dimorphic
T. equiperdum and the monomorphic strains which are generally
known by that name. It is equally impossible to determine whether
or not, in a country in which tsetse flies exist, the trypanosomes
causing the disease clinically known as dourine may not be capable
of transmission by these insects as well as mechanically in coitus.
It is, however, of interest to record the occurrence of a disease
clinically resembling acute dourine in mules in whom infection by
coitus may be excluded with certainty.
REFERENCES
Blacklock, B., and Yorki, W. (1913a). Trypanosoma vivax in Rabbits. Annals of Trop. Med.
and Parasitol.) Vol. VII, p. 563.
-(1913b). The Probable Identity of Trypanosoma congolense (Broden) and
T. nanum (Laveran). Annals of Trop. Med. and Parasitol Vol. VII, p. 603.
Bruce, D. (1911). Sleeping Sickness Bulletin , Vol. Ill, p. 422.
-(i 9 H)- Trans, of the Soc. of Trop. Med. and Hyg ., Vol. VIII, No. 1, p. 15.
Bruce, D., Hamerton, A. E., Bateman, H. R., Mackie, F. P., and Lady Bruce (1911).
Reports of the Sleeping Sickness Commission of the Royal Society, No. XI, p. 165.
Dutton, J. E., and Todd, J. L. (1903). First Report of the Trypanosomiasis Expedition to
Scnegambia (1902). Liverpool School of Tropical Medicine, Memoir XI.
Kinghorn, A., Yorke, W., and Lloyd, Ll. (1913). Annals of Trop. Med. and Parasitol .,
Vol. VII, p. 251.
Laveran, A., et Mesnil, F. (1912). Trypanosomes et Trypanosomiases. Masson et Cie., Paris.
Macfie, J. W. Scott (1913). Trypanosomiasis of Domestic Animals in Northern Nigeria.
Annals of Trop. Med. and Parasitol ., Vol. VII, p. 1.
Macfie, J. W. Scott, and Gallagher, G. H. (1914). Sleeping Sickness in the Eket District
of Nigeria. Annals of Trop. Med. and Parasitol ., Vol. VIII, p. 379.
Mesnil, F. (1915). Bull, de Vlnstit. Pasteur. Tome XIII, p. 248.
+92
EXPLANATION OF PLATES
Plate XXXV
Trypanosomiasis of mules resembling acute dourine.
Fig. i. Showing the advanced stage to which the disease had
progressed within twenty-four hours of the onset of the
symptoms.
Fig. 2. Showing the plaques on the skin.
Annals Prop. Med. & Parasitol., Vol. IX
PLATE XXXV
494
Plate XXXVI
Figs, i-14. A piroplasm of the brown rat —Nuttallia decumani ,
n. sp. x 2000.
Figs. 15-28. A small monomorphic trypanosome found in the
blood of a mare— T. congolense , var. equinum . x 2000.
Fig. 29. A red blood corpuscle containing an Anaplasma -like body
such as became very common in the blood of this mare in
the latter stages of the disease. X 2000.
*U
0
\
T
/
••
\
'
\
«
>
M •
%
S'
+ \
/ v •
*
/
/
V s
»•
>
s' ’
<
\
> •
ft)
vv\
■fr '
¥
!
i
/
0
\
/
/
4
•
S
rV *
\
-
.V,
1
}
0 ~
'
i
4
>
- i
• #
T\
•
<.■>,
V
u
M y
'<&
495
DIFFERENTIAL COUNTS AND THE
NEUTROPHILE BLOOD PICTURE OF
NATIVES—ADULTS AND CHILDREN—
OF NEW GUINEA
BY
A. BREINL
AND
H. PRIESTLEY
FROM THE AUSTRALIAN INSTITUTE OF TROPICAL MEDICINE, TOWNSVILLE
{Received jot publication 19 August, 1915)
INTRODUCTION
Our knowledge as to the blood conditions of coloured native
races is comparatively limited. It is only within the last few years
that Chamberlain and Vedder (1911) recorded observations on the
blood of Filipinos, Bahr (1912) on that of Fijians, and Marshall
and Meerwein (1913) on 'wild* natives of German New Guinea.
The Ameth blood picture has been studied in Filipinos by
Chamberlain and Vedder, and in New Guinea natives by Marshall
and Meerwein. Unfortunately only the observations on Filipinos
can be used for comparison, as our results, as well as those of
Chamberlain and Vedder, were obtained by the use of the original
Ameth classification, whilst Marshall and Meerwein employed a
somewhat different classification.
Chamberlain and Vedder pointed out that the Ameth index in
natives (Filipinos) showed a distinct increase when compared with
that of normal Europeans.
Our observations on the Ameth index of white school children,
bom and reared in the tropics (North Queensland), showed a change
in the same direction.
As opportunity offered, it was deemed of interest to investigate
the neutrophile blood picture of New Guinea natives, as well as the
percentages of the different types of white blood corpuscles, adults
and children being considered separately.
496
The material was collected during a journey through the coastal
parts of British New Guinea, blood films being made from natives
of different ages. Some selection was made in the case of adults,
blood films being obtained only from apparently healthy natives
who lived in comparatively clean villages, and all blood films
containing microfilariae or malarial parasites were discarded. As
the majority of the blood films were made during the day, infections
with microfilariae possessing a turnus could not be excluded. The
blood films were taken from natives who had been in contact with
Europeans for a considerable period, as well as from inhabitants of
villages which had not come under Government control, but no
striking differences could be observed.
With children it was practically impossible to make a similar
selection because of the wide distribution of yaws and malaria,
although children with apparent manifestations of yaws and skin
diseases were excluded, and films containing malarial parasites were
not considered in the present investigation.
The blood films were stained after Giemsa’s method, and for the
differential and Arneth counts the same technique was adopted as
employed in our blood examination of North Queensland school
children.
The results are based on the examination of 104 films from adults
and 50 from children, the latter, as far as one could judge, were
below 10 years of age.
DIFFERENTIAL COUNTS
For the differential counts 500 successive leucocytes were
enumerated.
Considerable variations were observed in the percentages of the
different types of leucocytes, if individual counts be considered
(see Table I). The average figure for the polymorphonuclear
leucocytes in adults was 5106 per cent., and the range lay between
28 9 per cent, and 81 per cent. This figure is somewhat below the
normal European average (55 to 70 per cent.) and is lower than the
average figure obtained for North Queensland school children
(5 it per cent.).
497
Table I.
Differential and Arneth Counts on the Blood of 104 Adult Natives from New Guinea.
Differential Counts per cent. Arneth Classification per cent.
Number
1
e-s
H
Is
2 1
A
!
Transitional
Large
Mononuclear
Lymphocytes
Eosinophiles
I
II
1
III
IV
v 1
1
1
71-2
4-6
J *4
13*4
9*4
4 2 ’5
38*0
!
i
' 7 -° !
2 *5
1
|
2
6i*6
3-6
3*4
28*8
2*6
3^-5
4 2 *5
19-5 1
5*5
1
3
6 i *4
3 **
2-2
25-6
7-6
40*0
46*0
I2 *5 '
*’5
_ 1
4
63-0
2-8
2*8
25-6
5-8
26*0
46-5
21*0
6-5
i
5 i
526
6-4
3-6
28-6
8-8
34 *o
50*0
-
2 *5
1*0
6 1
48*0
7 * 6
3 *o
33 *o
8-4
40-0
47*5
12*0
°*5
!
7 |
57 ' 8
46
0-2
21*6
> 5-8
17*0
38-0
35 *o 1
9 *o
'•° :
8
59 6 1
6-o
—
25-2
9-2
15-0
41*0
30*0 i
10*0
4 *°
9
58-8
6*o
2*0
26-8
6.4
33*5
41*0
20*0
5 *o
o *5
10
39 ' 6
4*4
5-6
368
136
34 *o
4 2 *5
20-0
3 -S
1
1
11 1
56*8 1
7-6
3 -o
2 7*4
5 * 2
34*5
44 *o
ITS '
1
3-5
o. 5 |
12
|
35 *°
3*3
i *4
2 9*3
31-0
34 *o
! 41*0
21*0
3*5
°*5
13 1
6o-8
7*4
2 *4
16-8
12-6
2 3 *° 1
| 44 *o
I * 6-5 ;
6*o
°*5 ;
-4 1
62-6
3-8
2*0 j
18*o
136
26-5
47*5
23-5
2 *5 !
—
i
r 5
49 *o
54
o-8
31-8
13*0
45-0
4**5
ID'S
3*0
1
■ 6 |
68-o
2*6
3 *o
* 7*4
9 -o
31*0
43 *o
22*0
4 -o
1
—
7 !
6o*o
2*4
o*8
18-8
18*0
25*0
46*0 i
2I *5 I
6*o
i *5
8
1
64*0
2-8
1*0
17-4
14*8
28*0
40-5
2 5‘5 !
5*5
o *5
9
40-0
4*8
2-8
29-2
2 3-2
26*0
38-5
26*5
8*o
1*0
0 1
59-8
2*0
3 *o ,
* 5 * 2
20*0
20*0
i
46*0
26-0
7 *o 1
1*0
1
46-4
3-6
3*2
30*6
16*2
39*5
i 1
!
41*5
1 3*5
4*5
10
2
43 * 2
4-6
2*0
37*4
12*8
40*0
4^*5
1 11*0
** i
3
32-8
5-6
>*4
33*4
26-8
36*5
44-0
! ,7 ’ 5
2*0
—
4
39*3
5 *o
i 1
27*2
27-0
39 *o
465
12*0
2 *5
— 1
5 1
76*0
3-2
!
1 1*0
, S -o
; 4-s
30*0
45 -o
21*0 !
3*5
0*5 i
6 -1
6o*8
1*6
! 2 ‘4
25-2
| 10*0
! 22 *5
, 4*5
2 5‘5 !
9*5
—
498
Table I.— Continued .
Differential Counts per cent. Arneth Classification per cent.
Number
Polymorpho¬
nuclear
n
c
c
n
H
Large
Mononuclear
0
'H.
E
Eosinophiles
I
II
III
IV
V
2 7
68-6
4 -o
1 -o
23*6
2*8
54*5
36-0
8-o
l '$
—
28
53*6
2*8
o-6
33'8
9*2
37-0
43-5
15-0
3*5
PO
29
5 2 * 2
5’5
3 ’°
26-8
,2 '5
2 7 * 5
48*0
* 9*5
4*5
°*5
30
627
3*3
22-0
io-5
40-0
445
i 3 *o
2*0
o-s
3 *
56-0
2 ’4
po
2 7*4
• 3-2
33 -°
45*5
.8-5
3-0
-
3 2
63-4
3-6
l-o
236
8-4
33 ’°
39 *°
22 '5
5*5
-
33
57-8
2 *4
i-6
30-2
8-o
.8-5
37 *°
26*0
7-0
i *5
34
53 -o
3*4
o-8
25-0
17-8
2 p 5
58-5
30-0
I o-o
-
35
63*0
2-8
0*2
23-0
I PO
22-5
46*5
20*0
ic-5
0*5
36
46-0
4 ‘ 2
i *4
38-8
9-6
29-5
37*5
23*0
9*0
PO
37
41-0
2-6
i*6
42-8
12*0
40*5
40-0
I 7 -0
2*0
°*5
00
ro
74 -o
2-8
1*0
1 5'4
6-8
20-5
39.0
2 9 -5
10*0
PO
39
5'*4
4*8
i-6
37-8
4*4
57-0
37 -o
5 *o
PO
-
40
41*6
2-0
PO
38-2
17-2
4P0
4 po
17-0
PO
-
4 *
50-2
2-0
3 *o
|
37 ’°
7*8
53 -°
37 -o
8*5
l *5
-
4 2
4*’4 !
i -4
; o*6
42-0
146
47-5
37-5
,2 ‘5
2 *5
-
43
40-0
i-8
1 12
5 PO
6-o
35 ’°
46-0
16*o
2-0
PO
44
8 po
1*2
o*6
i6-8
0-4
39 ’°
42-0
16*0
3 *°
-
45
40-0
2*0
l ,+
27-0
29-6
49 ‘°
4 2 *5
8-o
o *5
-
46
60*2
2-8
i 14
25-0
io*6
35*5
5°*5
ip 5
2*0
o-s
47
59 *o
i *4 |
1 ** 2
30-8
7-6
2 PO
41*5
26*0
I PO
o *5
48
43*6
3*4
p8
38*4
12-8
29*0
50-5
16*o
4*5
-
49
66-2
2*0
o -4
24-8
6-6
39’5
40-0
18-o
2*0
o-s
5 °
51-0
i-6
1 *o
37-6
8-8
4°*5
43 -o
I2 ’5
3*5
o *5
5 1 ;
44 -o
1*2
o-8
37-0
17-0
25-0
47*5
2PO
4 *o
2*5
5 * !
396
4 -o 1
2 *4
44 -o
10*0
4P0
43 *°
12*0
4 *°
-
499
Table I.— Continued.
Differential Coimts per cent. Amcth Classification per cent.
Number 1
Polymorpho¬
nuclear
Transitional
Large
Mononuclear
Lymphocytes
Eosinophiles
-
I
II
III
IV
V
53
t
j 5°'4
i-6
1*0
34 *o
i
13*0 ,
37*5
36*5
22*5
3 -o
o -5
54
| 6i*2
2 ’4
1*2
27*0
8*2
35 *o
39 *°
21*5
4*5
—
55
41*6
4*4
3 -o
4 i *4
9-6
38-5
50-5
10*0
1*0
—
5 «
28*9
2-6
i*6
47-8
i 9 ** ;
4 2 *5
43-0
13*5
1*0
—
57
45-8
i-6
2*4
40*8
9-4 |
20*0
49.0
2 4*5
5-5
i-o
5*
45 °
2 *4
2*0
35-6
15-0
* 4*5
41*0
29*5
,2 *5
2 *5
59
49*4
24
2*0
32*6
136 !
26-5
47 *°
24*0
2 '5
—
60
1
60-4
1*0
! *4
19-4
17*8
22 *5
47 *°
26*5
4 -o
—
61
49*4
2*2
1*2
33 * 2
14-0 .
37*5
455
14*5
2*0
o *5
62
48-4
2-4
2*2
38.2
8*8
27*5
46*0
22*0
4-0
o -5
63
47 * 2
1*8
2*0
34-6
.
14*4
27*0
41*0
29*0
3.0
—
64
52*2 1
I *2
o*8
'
34-4
■■•4 |
27*5
47 *° |
2 3*5
1
i *5 j
i
°*5
65
5 6 * 2 1
2-0
2-8
23*2
.5-8 !
3 2 ‘5
45*5 ;
20*5
" 5 !
—
66
44*4 i
1 *4
i*8
40-6
11-8
*8*5
1
42*0 1
28*5
1 10*0
1
1*0
6 7
72-6
**4
i*8
.
21*4
i*8 i
33*5
5 i *5 1
13-5
i 1*5 !
_
68
56-8
24
1*0
1
; 2 7 *°
12*8 1
18*5
46*5
2 4*5
9*5 |
1*0
69
39 * 2
2 *5
i*6
1
1 36*5
20*2
24*0
44*5 |
28*0
3*5
l
70
48-6
2*0
o*6
1 3 2 * 2
16*6
2 7*5
48*0
,
2I *5
! 3-o
—
7 *
48-0
1*2
2*2
i 4 1 * 2
7*4
24*0
47 *o
1 2 4*5
j
1 3*5
1*0
7 2 !
47*2
2*6
1*2
28*2
20*8
18*o
43*5
i 3 i *5
1
7 *o
—
73 j
6i*o
1*4
2*8
26*0
8*8
■ 7 -S
44*5
3 i *5
6*o
o *5
74
51-6
i-8
1 2*2
34-8
9*6
19-5
46*0
30-0
■
4*5
75 j
55.0
2*6
1*6
I f
j 2 9*4
n *4
.5-0
45 *°
30*0
1 9 *o
1*0
76
46*2
2*0
| 2*2
! 3«-6
11*0
19*0
45*5
30*0
1
5 *o
1
0-5
77
45.0
2-8
2*2
! 4°'4
9*6
33*5
44.0
19*5
2-5
o *5
78
56-2
2*2
i 2-4
j 3 2 '8
6 *4
i
34 *o
46*5
17*5
1 2-0
1
—
5 01
The average number of lymphocytes (32* 1 per cent.) was definitely
increased, and the percentages in the individual counts varied
between 13*4 and 53*3 per cent. These observations are in agree¬
ment with those of previous investigators, namely, that the relative
number of lymphocytes in the blood of native races in the tropics is
definitely increased.
The relative number of the eosinophile leucocytes was larger than
in Europeans; the average was 11 *97 per cent, and the range was
between 04 and 31 per cent.; 57 of the counts were above 10 per
cent.
It is impossible to determine how far this eosinophilia can be
accounted for by helminthic infections, since examinations of the
faeces for the presence of intestinal parasites were impossible; but
Bahr has pointed out that eosinophilia was well marked in the blood
of Fijians, who showed neither ova of intestinal parasites in the
stools nor microfilariae in the blood. Eosinophilia, as is well
known, may be due to other causes than infections with helminths,
such as skin diseases and the presence of ectoparasites, both fairly
common amongst the natives.
The relative numbers of transitional and large mononuclear
leucocytes corresponded on the whole to figures considered normal
for Europeans.
The differential counts of 50 children (see Table II) showed
similar but more pronounced changes. The average number of
polymorphonuclear leucocytes was 4006 per cent., with a variation
in the individual counts between 18*4 and 65 per cent.
There were on the average 42*86 per cent, lymphocytes, and no
count showed less than 29 per cent.
The average number of eosinophile leucocytes was increased
(13*24 per cent.), the individual counts ranging between 1 and 42*4
per cent.
There was no alteration in the relative number of large mono¬
nuclear and transitional leucocytes.
502
Tabu II.
Differential and Ameth Counts on the Blood of 50 Native Children from New Guinea.
i
Differential Counts per cent.
Ameth Classification per cent.
j
u ,
0
M u
D.
1
O
u 1
n 1
jj 1
t> *o 1
00 3
1
tn \
O
>N I
u
0 1
i
cu
1
1
|
K
JU
C
•a 1
3
2
1
e w
Is
<2
•s !
c
rt
1-1
*1 0
^ C
*2 1
^ 1
-C
O,
1. I
J j
0
c
•a
W
I
i
I
II
III
iv !
1
V
X
*»
V
c
<
I
40*2
o-8 '
1
o-8
52.4 1
5-8
44 -° !
42-0
13*0
1*0
—
Z
41*0
o-8
i-8 1
1
5 '-+ i
5 *o
31-0 ,
I
46-0
20*0
3 -° |
-
3
30-9
c 9
°*9
61 *9
5*4
37-5 ,
40-5
20*0
2*0 |
—
4
5 2-2
2*6
i *4
38-4 1
5*4
2 , 5 |
42*0
26-5
8*o j
2*0
5
37-6
1*1
«*3 I
47-6
12-4
4**5
43 -o
! 14*0
1
i *5
-
6
32*3
i *7
, 4 ;
53 *i
ii *5
40*0 *
44.0
1 14-5
'‘ 5 1
—
7
364
i -4
o' 9
54-6
67
39*5 |
35 -o
1
; 22 *5
I |
3 . 0 .
-
8
26*9
i *5
o -9
59*5
11*2
48*0
40*0
12-0
—
-
9
40*8
3-8
" 4
30-0
24*0
28-5
32-5
25*0
ii *5
2*5
IO
3 i*o
3*6
2-2
32-6 ;
3°*6
32*0 1
40*0
! 21*0
1 6-s
o -5
11
37-5
2*0
0-5
367
23*3
29*0
40*0
2 3*5
7 *o
o *5
12
28-6
4-1
0-7
34 -°
32-6
33*5
43 *o
1 19*0
3*5
1*0
1
1
*3
46-6
I
4-0
i 6-6
39 *o
3*8
44*5 ,
4 i *5
1
120
2*0
—
1
1
i 4
50-0
4-3
2* 1
36*0
7*6
43*0
44*5
12*0
0*5
-
1
j
15
52*6
4-6
i*6
34 -o
7*2
j 6i*o
36*0
3 *o
—
-
!
16
39 -o
-6
2*0
434
14-0
| 35*5
46-5
i 4*5
3*5
-
*7
46*0
I 2-8
1
i-6
38-2
1 1*4
j 38*°
37 *°
19-0
5*5
o *5
1
18
33*3
j i *5
0*2
49-2
15*8
i 49 *o
39*5
9 *o
2*5
-
1
19
i 27-9
I
J 2 ‘4
i *5
50*6
17*6
34*5
48*0
1 16-5
1*0
-
20
1 38-3
37
1*0
4 i *3
157
! 35 -o
46-5
15*5
2*5
o -5
21
51-4
, 2-6
o-6
35*2
10*2
; 6-5
3 i *5
7 *o
—
-
22
28-6
2*0
2-5
39*2
2 7*7
! 44-5
43*5
n *5
o -5
-
^3
! »7
2*0
°*3
53*3
217
! 32*0
48*0
.8-5
i *5
-
24
47-6
! 1*0
2*0
1 42*8
6-6
53 *o
39*5
6-5
1
1*0
-
25
4 '-*
2*2
2-8
48*6
5*2
! 44 *°
1
1 48-5
7-5
l
—
503
Table II.— Continued.
Differential Counts per cent. Arneth Classification per cent.
Number
Polymorpho¬
nuclear
Transitional
Large
Mononuclear
Lymphocytes
Eosinophiles
I
II
III
IV
1
V
Arneth Index
2 6
t 40-6
i *4
i-8
44-6
ii*6
47*5
42*5
9*0
1*0
—
27
oc
vO
r*1
! *5
i *5
42*4
17*8
39*5
+6-;
13*0
1*0
—
28
' 35*3
i *3
2*2
547
6-5
42*0
465
11*0
°*5
—
29
53*4
2*2
1-4
32-6
io*4
29*5
48*0
20*0
2*5
—
30
37*3
1*1
i -4
47 -i
13*1
48*0
40*0
12*0
—
—
3 *
, 41*8
1
*•4
i-6
45-8
9*4 ,
24*5
44‘5
25*5
5*5
—
32
3*7
*•3
o-8
54*2
11*0
30*5
465
21*5
*‘5
—
33
28-9
|
o -9
1*0
56*1
* 3 -i 1
39-5
44 *o
■ 4-5
2*0
34
40-2
2*2
o*8
31-2
256
46*0
40*5
12*0
f -5
35
! 25-0
4-8
3 ‘*
55*4
"7 !
■
57 *°
34 *o
8*0
1*0
—
36
35-3
2-0
*•4
39-5
21-8 ]
55-5
37-0
6*o
p 5
37
+ 5-4
i-6
! *4
40*2
u *4
5 ,- °
33-5
* 4*5
o *5
i
, °*5
38
43 *o
3*2
38-2
15-6
5 2 '5
36*0 j
:
10*5
1*0
| —
39
41-2
4-0
1-2
42-0
11*6
36*0
45 *°
16*5
2*5
40
54-8
3-8
—
37*4
4 '° ,
45 ‘° I
1
45 *° 1
8-5
*•5
4 i
18*4
i -3 1
—
37*9 !
4 2 ’4 1
49.0
40*0
8*o
3 *o
■
—
42
00
6
1-3 ;
o -5
48-4 |
11 -8 1
42*0
42*5
12*5
2*5
o *5
43
45*3
8-8 1
1*0
00
62 ;
i j
4 1 “5
37*5
1 7’5
3-0
o *5
44
37 -i
3 -o
>*5
38-8
19*6
6o*o
35 *°
4 *o
1*0
—
45
40-4
2*6
—
38 -*
• 18*8
42*5
1
44 *o
13*0
o *5
—
46
59 -o
3*8
i-8 j
30*6
4-8
44 *o
42*0
12*0
2*0
—
47
65*0
4*2
o-8
29-0
, ‘° 1
58*0
35 *o
7*0
—
—
48
53*2
3 -o
i *4 J
35*4
7 *°
56*5
36-5
1
7 *o
—
1
49
45*4
2-4
1
1*2
49*4
i*6
60*5
1 29*5 |
9*5
I
—
°*5
1
50 1
49-0
i-6
1-2
32*4
15*8
48-5
i 1
1 35 -o
* 3-5
2*0
1 *o
Average
40-06
2*47
1-36
42*86
* 3-24
42*96
, 40*9°
; 13*80
2*13
, 0*21
83*86
5°4
ARNETH COUNTS
For the Arneth count 200 consecutive white cells were enumerated
in two separate lots of 100, and only those counts were considered
where there was close agreement between the two series.
The average for the Arneth index (the sum of leucocytes of
classes I and II) in 104 adult natives was 74 (see Table I). A
comparison with the average index of Europeans (40) shows that the
index is considerably increased in adult New Guinea natives. It
is practically the same as that in North Queensland school children
(74*5), and is higher than that found by Chamberlain and Vedder
in their examination of 50 Filipinos (65 8).
The average Arneth index of 50 New Guinea children (1 to 10
years old) was still higher, being 83 86 per cent, (see Table II).
This close correspondence between the Arneth index of adult
New Guinea natives and that of North Queensland school children
indicates that the increase in natives cannot be attributed to infection
only, since the school children examined were, as far as one could
judge, perfectly healthy.
The comparative increase in the Arneth index of native children,
when compared with adults, probably finds its explanation in the
greater incidence of active and latent infection amongst native
children. Yaws and malaria were found endemic in the majority of
the coastal villages of British New Guinea, and although children
who were obviously suffering from yaws, and those who had malarial
parasites were excluded, it is probable that the majority of the
children were, or had been, suffering from either malaria or yaws,
or most likely both.
This assumption is, moreover, strengthened by the observations
of Scott Macfie on the Arneth counts of cases of malaria in West
Africa. The blood of such cases showed an increased Arneth
index, which persisted even after the disappearance of the parasites
from the blood and the recovery of the patient.
CONCLUSIONS
I. Differential counts on 104 adult natives of New Guinea show
a decrease in the number of neutrophile leucocytes and an increase
in the number of lymphocytes and eosinophiles.
SOS
2. Differential counts of 50 native children show similar but
more pronounced changes.
3. Ameth counts of adult natives show a marked shift to the
left, the average Ameth index being 74 0.
4. Ameth counts of 50 children gave an average Ameth index
of 83 86.
5. The identity of the Ameth index of adult natives with that
of healthy school children in North Queensland strengthens the
assumption that climatic influences, per se 9 cause a ‘shift to the
left *; but the still greater shift in native children is probably due to
active or latent infections.
REFERENCES
Bahr (1912). Filariasis and Elephantiasis in Fiji. Supplement No. i, Journal of the London
School of Tropical Medicine, p. 92.
Breinl and Priestlky (1914). Changes in the Neutrophile Blood picture of Ameth observed
in children living in Tropical Queensland. Annals of Tropical Medicine and Parasitology,
Vol. VIII, p. 565.
Chamberlain and Veddir (1911). The study of Ameth’s nuclear classification of neutrophiles
in healthy adult males and the influence thereon of race, complexion and tropical
residence. Philippine Journal of Science , Vol. VI, p. 403.
Marshall and Meerwein (1^13). Ueber das leukocytare Blutbild, einschliesslich Verschiebung
der Neutrophilen bei wilden Eingeborenen von Neuguinea. Folia Haematologica,
Bd. XV, p. 229.
Scott Macfie (1915). The significance of Nuclear variation of Neutrophile Leucocytes (Ameth
counts) in W'est Africa. Lancet, Vol. 1, No. XVIII, p. 911.
A CASE OF DYSENTERY IN A MONKEY,
IN WHICH AMOEBAE AND SPIRO-
CHAETES WERE FOUND
BY
J. W. SCOTT MACFIE, D.Sc., M.B., Ch.B.
WEST AFRICAN MEDICAL STAFF
(Received for publication 27 September , 1915)
Plate XXXVII
INTRODUCTION
A small monkey, Cercopithecus petaurista , in which a strain of
the human trypanosome had been sent to Accra, Gold Coast Colony,
from Sunyani in June, died unexpectedly on August 5th, the
sixty-fifth day after inoculation. The monkey had been observed
to be suffering from diarrhoea for some time, but as it appeared to
be healthy otherwise, little attention was paid to this fact.
At the autopsy the body was found to be somewhat wasted, and
all the organs and tissues were decidedly anaemic. The lungs
appeared to be healthy. The heart was pale and flabby, and the
blood was unusually thin. In the abdominal cavity there was no
excess of peritoneal fluid; the spleen was considerably enlarged but
the other organs were healthy looking, but anaemic. There was no
hypertrophy of the lymph glands. No parasites were found in the
gut, but throughout its length the large intestine was congested and
studded with ulcers. These ulcers were most numerous near the
caecum, but were scattered all over the mucous surface down to the
rectum. Each ulcer was roughly circular, with a central slough, and
raised, ragged, blood-stained edges (see Plate XXXVII, fig. 12).
In the blood trypanosomes were found, but they were not
numerous. During the forty days the monkey had been under
observation at Accra the trypanosomes had always been scanty or
rare in the blood, and no symptoms of trypanosomiasis had
manifested themselves. It is improbable that the death of the
508
monkey was due to trypanosomiasis. The short duration, sixty-five
days since inoculation, the absence of all clinical symptoms, and of
enlargement of the lymph glands, and the fact that another small
monkey inoculated at the same time is still alive and well although
more heavily infected, all tend to prove that death must have been
due to some intercurrent disease. The extensive ulceration of the
large intestine suggests itself as this cause, and I believe that death
was due to amoebic dysentery.
AMOEBAE FOUND IN THE LARGE INTESTINE
In fresh preparations of the contents of the large intestine
numerous amoebae were seen. They were very active, extruding
clear pseudopodia in all directions from their surfaces, but did not
progress rapidly across the field of the microscope. In each a single
nucleus was visible through the granular and vacuolated endoplasm,
and in some cases red blood corpuscles were seen to have been
ingested. There was no contractile vacuole. When stained the
amoebae measured from 12// to 30 n in diameter (Plate XXXVII,
figs. 1-6). The cytoplasm was much vacuolated and contained
numerous bacteria-like chromidia. The nucleus was somewhat
indistinctly stained by Leishman’s method, but appeared to possess
a fine chromatin network, but no distinct karyosome. In considering
this fact it should be remembered that the specimens were obtained
at an acute stage of the infection. The commonest forms of the
amoebae measured 12/x to 15/i in diameter; but a few much larger
individuals were found which measured 26 ju to 30/x (fig. 3). The
latter forms, besides being vacuolated, often contained a
number of ingested red corpuscles, and in some cases this
may have accounted for their large size. It is possible, however,
that the large amoebae might have been of a different species; but
as some forms of intermediate size were also present, it seems more
probable that all the forms were stages in the life cycle of the same
parasite. Numerous cysts (figs. 7-10) were also present measuring
as a rule 12/x to 18// in diameter, but some larger forms were seen
which measured as much as 33/1 . These cysts had a thick wall,
which stained red with Leishman’s stain, and a single large
(? glycogen) vacuole occupying the greater part of the interior. The
vacuole stained a pink colour. In the narrow band of cytoplasm
the nuclei were visible, as oval or rounded bodies with a palely
stained chromatin network. The number of nuclei varied greatly;
in some cysts there was only one, but as a rule there were several.
In one cyst eight nuclei were counted (fig. io). In sections
large numbers of amoebae were seen in the bases of the ulcers and
penetrating into the adjacent layers of the intestine.
The cysts were in some respects unlike amoebic cysts, and
resembled superficially the bodies called ‘cysts of Trichomonas
intestinalis 9 which Alexeieff (1911) considers to be ‘en r6alite un
Ascomycete voisin des Levures/ and for which he has proposed the
name Blastocystis enterocola. If Alexeieff is correct in his interpre¬
tation, it is possible that the cysts found in this monkey at Accra
may have been of a similar nature. It should be mentioned,
however, that the characteristic nuclear structure described by
Alexeieff, 1 une calotte chromatique peripherique separee par un
halo clair du reste de la substance chromatique finement granuleuse,’
was not observed; and no indications were seen of the two forms of
division traced by him, namely, that by budding (‘ bourgeonne-
ment’), and that ‘par etranglement (division plasmotomique)/
Neither did the cysts appear to be enclosed in a mucilaginous
covering as Blastocystis enterocola apparently is. It seems more
probable therefore that, in the case of this monkey, the cysts should
be associated with the amoebae that were so numerous in its
intestine.
Amoebae have previously been described from various monkeys,
Mathis (1913) described two types of cysts found in healthy monkeys
(Macacus rhesus and Macacus tcheliensis ) in Tonkin, and gave a
r6sume of the literature on this subject and enumerated the
Entamoebae found by various authors in monkeys. None of these
parasites seems to have resembled those found in this case.
Swellengrebel (1914) described by the name Entamoeba chattoni
an amoeba found in Macacus rhesus at Deli. This parasite
measured in the amoeboid form between 13 n by 12 n and 12 /*
by 9/1, and in the cystic 9/1 or 8/1 in diameter. The cysts were
described as containing a glycogen vacuole, but only uninucleate
and binucleate forms were observed. A comparison between the
illustrations given by Swellengrebel, and those on the plate attached
5io
to this note, will show, I think, that the amoeba from Macacus rhesus
differed from that found in Cercopithecus petaurista . In the
majority of cases the parasites have been found in healthy animals,
and their pathogenic nature is doubtful. I have not been able to
find any previous description of a case of amoebic dysentery in a
monkey ( Cercopithecus ) similar to the above, and I would therefore
suggest the name Entamoeba cercopitheci for the parasite found in
this case.
A SPIROCHAETE RESEMBLING SPIROCHAETA EURYQYRATA
FOUND IN THE LARGE INTESTINE OF THE SAME MONKEY
In addition to the amoebae, vast numbers of minute spirochaetes
were seen in the smears made of the contents of the large intestine
and rectum of this monkey (fig. n). These spirochaetes were
extremely slender, and had finely-pointed extremities. They
stained uniformly a reddish colour with Leishman’s stain. When
living they were very active, but owing to their small size were
difficult to study. Twenty-five individuals, taken as they came,
were drawn with a camera lucida and measured by the tangent line
method. The average length was 5*28/1, and the range from
4/1 to 7/4. Some of the spirochaetes consisted of a single loop or
spiral, others showed two and a half such coils, but the majority
showed one and a half or two, and measured 5/4 to 6/4 in length
(see Table 1).
Table I. —Measurements of twenty-five specimens of a spirochaete found in the large
intestine of a monkey.
Length in microns
Number measured
Average number of loops or
spirals
4
5
**3
5
10
i-6
6
8
1-87
7
2
225
The morphology of these spirochaetes closely resembled that of
Spirochaeta eurygyrata , a species found by Werner (1909) in his own
faeces at a time when he was apparently in good health. Similar
spirochaetes have been found by J. G. and D. Thomson (1914) in
the stools of apparently healthy individuals; and, at Accra, I have
found what would appear to be the same parasites in enormous
numbers in the faeces of two patients who were suffering from
diarrhoeic symptoms. Spirochaeta eurygyrata is said to measure
4*6/* to 7 3 fi in length, with usually two curves, and to possess a
very flexible body; a description that would apply very well to this
spirochaete from Cercopithecus petaurista , and it would, no doubt,
be inadvisable to give the latter parasite a specific name merely
because it occurred in a different host.
There was no evidence in this case that the spirochaetes were
pathogenic, and it is probable that they were comparable with the
small spirochaetes frequently found in the gut of animals, and man,
in West Africa. It is not improbable, however, that the debilitated
condition of the host, and the diseased state of the large intestine,
may have provided the favourable medium which allowed these
organisms to multiply; and that the vast number of spirochaetes
thus engendered may have contributed to the ill-health of the animal
which finally culminated in its death.
REFERENCES
ALixiiKFr, A. (1911). Sur la nature des formation* dites 1 Kyste* de Trichomonas intcstinalis.*
C. R. Soc. Biol.j Tome LXXI, p. 296.
Mathis, C. (1913). Entamibe* de* Singe*. Bull. Soc. Mid.-Cbiruro. de Vlndocbine. Vol. IV,
p. 388.
SWELLKNGRIBIL, N. H. (1914). Dierlijke Entamoeben uit Deli. Geneesk. Tijdscbr. v.
Nederl. Indie . Vol. LIV, p. 420.
Thomson, J. G. and D. (1914). Some Researches on Spirochaetes occurring in the
Alimentary Tract of Man and some of the Lower Animals. Proc. Roy. Soc. of Med.
Vol VII, p. 45.
Wsrnka, H. (1909). Ueber Befunde von Darmspirochaten beim Menschen. Centrabl. f.
Baku Abt. 1, Orig. LII, p. 241.
512
EXPLANATION OF PLATE XXXVII
Figs. i-io. Entamoeba cercopitheci , n.sp., from a monkey
(Cercopithecus petaurista ) at Accra, Gold Coast, West
Africa. Figs. 1-6, vegetative forms; figs. 7-10, cysts,
x 1000.
Fig. 11 A spirochaete of the type of Spirochaeta enrygyrata from
the large intestine of the same monkey. x 2000.
Fig. 12. A piece ot the large intestine of the monkey showing the
ulcers, | natural size.
Annuls Trap. Med. &• Parasitology, Vol. IX.
PLATE XXXVII.
AMOBAE AND SPIROCHAETES,
FROM THE GUT OF A MONKEY.
PRELIMINARY NOTE ON THE
GENERAL DISTRIBUTION OF GLOSSINA
PALPALIS, Rob-Desv., IN THE DISTRICT
OF LOMAMI, BELGIAN CONGO
BY
Dr. J. SCHWETZ
mIdECIN CHEF DE SERVICE, CONGO BELGE
(Received for publication 12 May , 1915)
With Map
Three principal factors determine the distribution of tsetse-flies
in any region; first the climate; secondly the vegetation, whether
forest, ‘park/ ‘bush/ savannah, or steppes; and thirdly, the distri¬
bution of water, such as lakes, rivers, streams and marshes. Upon
the variable combination of these factors depend not only the
presence or absence of all tsetses, but also of particular species or
groups. Knowing then the three factors in question for a Central
African region, it is possible to foresee and to predict the presence
or absence of one or other group of tsetses.
The district of Lomami occupies the region situated between
5° and 9 0 S. latitude and between 23 0 and 26° E. longitude. Thus
it is a tropical country with two seasons more or less marked
according to the latitude. The altitude varies from 1,000 metres in
the South to about 500 metres in the North. If from an administra¬
tive point of view Lomami belongs to Katanga, it is separated both
geologically and botanically therefrom. Here it would be out of place
to discuss the geological differences, notably the absence of mines,
but I would recall that, as regards vegetation, Katanga is
characterised by an almost continuous ‘ park * (or bush as it is termed
in Katanga) which is interrupted only by grassy plains (savannah or
steppes) on the higher plateaux. Only the S.E. border of the
Lomami district partakes of the park-like nature of Katanga,
the remainder being occupied by more or less typical savannah
5*4
intersected here and there by varying sized belts, along the rivers,
and by stretches, in the lower parts, of true equatorial forest.
However, in certain regions (for example between Mutombo-Mukulu
and Kabongo) areas recalling the ‘ park * are found, but these are
isolated and do not form large continuous stretches as in Katanga
proper.
The terminology used in connection with these different types of
vegetation is very embarrassing and often leads to confusion. Each
author interprets the various terms in his own fashion, and in
certain publications the explanations of 1 bush 9 (brousse), ‘ wooded
savannah* (savane boisee) are so vague that it is impossible to
understand them. This is chiefly due to the fact that nature is
infinite in its variations and lends itself by no means easily to our
schematic definitions. Such schemes are, however, useful in
facilitating the comprehension of the phenomena and thus in
systematising our ideas. Therefore, I am constrained to give my
own interpretation of the different names applied to the various
forms of tropico-equatorial vegetation. First, I would mention that
the word ‘ bush * does not define any particular type of country; in
African usage it indicates any area outside the station, and is
therefore a general term with a somewhat negative significance.
There is little need to give any definition of the term equatorial
forest. Everyone knows that it is a collection of very tall trees
whose upper branches give continuous and permanent shade and
whose trunks are covered with lianas and creepers and surrounded
with bushes and small trees, forming an inextricable and impassable
barrier. In the regions of the savannah, at least in Lomami, the
forest is encountered in the lower swampy parts and along the rivers,
forming the so-called galeries boisees along the latter. By the term
‘ park 9 (‘ pare ’) is meant an area covered with trees, usually stunted
and deformed, which provides but little shade; lianas and creepers
are almost entirely absent, and travelling between the trees is easily
accomplished. The savannah (savane ) is a flat or undulating stretch
of country covered with grass; if, however, the grass is short the
term steppe is usually applied. The savannah is rarely covered
with grass alone; as a rule, groups of bushes, small trees and shrubs
occur, and sometimes these bushes, etc., are numerous and fairly
tall, in which case the term wooded savannah (.savane boisie) or
S l 5
bush (brousse ) is used. The wooded savannah thus represents a
type intermediate between the true savannah and the park.
It will be remembered that all tsetse-flies require shade. While
G. morsitans has need of a certain freshness and does not require the
immediate proximity of water, G. palpalis must have a warm and
damp atmosphere, and the immediate presence of water in some
form—lakes, rivers, swamps, or, at least, very small marshes.
G . morsitans , in fact, exclusively inhabits the ‘park ,* while
G . palpalis frequents exclusively the immediate neighbourhood of
water surrounded—or even covered, as in marshland—with dense
vegetation. Therefore, in all stretches of true equatorial forest
G. palpalis may be everywhere, while in the savannah regions it is
able to exist in the galeries bois6es along the water courses, and in
the lower wooded parts. The savannah itself is always exempt from
tsetse-flies . G. morsitans is to be found neither on the plains nor on
the plateaux, and G. palpalis does not occur in the neighbourhood
of water when the latter is bordered only with grass, reeds or
papyrus.
Following the above explanatory remarks it will be sufficient to
add for the preliminary comprehension of the general distribution
of G. palpalis in the Lomami district, that the whole region is
intersected with thousands of wooded watercourses and is covered
with numerous lakes, lagoons and marshes of varying size, all of
which are also often wooded. In certain regions of Africa, notably
Katanga, G. morsitans is, among other things, characterised by its
ubiquity, while in Lomami G. palpalis occurs only locally.
Almost all my ideas regarding the habitats of this tsetse-fly—
ideas which I had acquired during my first stay in the Congo,
notably at Tanganyika—have been upset after several months*
travelling in Lomami. Certainly the general and well-known
principle, that the existence of G. palpalis depends upon two
fundamental conditions, viz., the presence of water and shade-giving
vegetation, remains, and will remain, unaffected, but a principle of
so general a nature is only sufficient for amateurs. Formerly I had
elaborated certain fundamental principles which enabled me to
predict, on approaching a place, whether G. palpalis would be
present or not—and I was seldom mistaken. Here, however, the
principles upon which I relied seemed to collapse entirely. For
example, I held previously that all small rivers or streams were
exempt from G. palpalis , and now I found it often occurred on quite
insignificant watercourses. Further I had stated that in very
marshy places there were no G. palpalis (and this fact has been
verified by numerous observers in East Africa), but here I discovered
it in great numbers in the deep marshes and on the swampy papyrus-
covered borders of rivers. Finally, to my great amazement, I was
also attacked by G. palpalis among certain very small and
insignificant swamps. But should we infer that the habitat of this
tsetse-fly is totally different in different regions? By no means—
it is merely that certain adaptations to local conditions occur which
require analysis and interpretation.
It may be of interest if I discuss a few of the peculiar cases which
have thus disconcerted me.
I. Lake Boy a (or Morid)
About two hours* journey from Kabongo is a small lake, more or
less ovoid in form and approximately 4 to 6 kilometres in diameter.
This lake is situated in a swampy hollow, and the approach to it is
rendered very difficult, and in many places absolutely impossible.
It seems that until quite recently (about three years ago) there
existed a true lacustral village built on piles, but now there remain
only three small fishing villages situated near the lake. Leaving one
of these villages in order to make a short tour of the lake in a native
boat, I walked in the forest for a time and suddenly arrived at an
abrupt bank, sharply dividing the forest from a marsh covered with
papyrus and other plants. I descended to the marsh, a difference in
level of several metres, and preceded by some natives who pointed
out the safe spots on which to tread, commenced to flounder in an
evil-smelling slime. The marsh presented the appearance of a series
of small islands of papyrus and other plants, disseminated on the
surface of a gelatinous substance of a semi-solid or semi-liquid
nature. The surface of this mass was for the most part bound
together by the superficial roots of the vegetation, where this
happened to exist, but the further one progressed the more difficult
and perilous became the advance, and one was in danger every
moment of being engulfed. Ultimately a narrow channel, made by the
natives, was reached, and a pirogue or native boat obtained. This
5i7
canal enables the natives to traverse the intermediate zone between
the lake proper and the surrounding marsh. In reality there is no
distinct limit between the lake itself and this intermediate semi-liquid
zone; the latter gradually becomes more and more liquid the nearer
one approaches the lake. A glance at the surroundings was almost
sufficient to convince me that Lake Boya was exempt from Glossina
palpalis. But while standing on the edge of the channel awaiting
the return of the natives, who had gone to look for a second boat kept
in a neighbouring channel, I perceived to my great astonishment that
a specimen of G. palpalis had settled on myself—shortly afterwards
also I noticed a second example on a native. During my tour of
some hours on the lake, in which both the centre and the banks were
visited, no further specimens were seen in spite of most careful
search; neither were any visible during my subsequent return on foot
across the marsh. When, however, I approached the steep bank at
the edge of the forest belt, I suddenly perceived several tsetses, and
later found that they were very numerous in the forest itself. The
explanation of the appearance on the marsh of the two flies was thus
provided. On the immediate borders of the lake, that is, in the
marsh covered only with plants—papyrus and occasional reeds—
there are no flies whatever. This marsh, however, replaces the
reservoir of water required, and when, as in the proximity of the
forest belt, shade-giving vegetation is present, the conditions
necessary for the existence of this species are supplied. The
apparent absence of the fly from the forest region during my first
incursion on approaching the lake was undoubtedly due to the
earliness of the hour. The two flies encountered in the marsh
therefore had evidently followed me and my black companions from
the forest.
2. River Kekey
This river, a left affluent of the River Lomami, presents during
the greater part of its course special characters recalling those of
Lake Boya. In the neighbourhood of the villages Mulenda and
Inga (Kabinda-Kisengwa route) the river itself is not visible; a long
valley about one kilometre wide with distinct, more or less elevated
banks can be seen. This valley is in reality a deep marsh covered
with grass, reeds and papyrus, and bordered on each side at
5 i8
intervals with trees of various sizes, bushes, etc. Within this valley,
sometimes in the centre, sometimes near one or other side, winds the
Kekey river—a river of some little importance. Near the village
Mulenda, where the ‘ marsh-valley * is very deep and dangerous, is
an artificial canal commencing at the border of the valley itself, and
used by the natives for fishing purposes. As it happens, the marsh
is very rich in a species of eel, which constitutes an important branch
of the local commerce. The river also supplies a species of trout.
G. palpalis is more or less abundant on the edges of this marsh.
But another question presents itself: is the fly to be found on the
banks of the river itself, that is in the centre of the marsh ? This is
without cover, there are no trees or bushes, and theoretically the fly
ought not to be present, but as a native boat was unobtainable in
order to reach the river via the canal, I was forced to forego personal
observation. Judging by what I have seen since under absolutely
identical conditions on the River Lomami, I conclude that G . palpalis
is not in evidence on the River Kekey itself.
3. River Lomami
This river is of great length, and traverses successively, wooded,
grassy, forestal, mountainous, swampy and open flat regions;
accordingly it presents much variation in the formation of its banks.
In the region of the 5th parallel the Lomami runs between two walls
of forest, while in the neighbourhood of the 7th parallel the banks of
the Lomami are in the form of papyrus-covered swamps recalling
those of the River Kekey* At a certain distance from the river a
chain of more or less wooded hills is visible on each side. In places,
however, these hills approach quite close to the river, receding again
later to give place to the marsh.
No single specimen of G. palpalis was seen so long as the
river flowed between the swampy banks, but directly the course
approached woody and hilly borders, the fly appeared and continued
in evidence until the marshy, papyrus region was re-entered.
In all the above places then—Lake Boya, the River Kekey and
the River Lomami—the same thing occurred. The marshy,
uncovered banks, whether of lake or river, are exempt from
G. palpalis , but such marshes are the equivalent of open water,
and, therefore, when bordered with shade-giving vegetation, the
conditions necessary for the existence of the fly are present.
S l 9
About one hour from Thielen St. Jacques (in the Kanda-Kanda
territory) is a large village Tshipama. Five minutes’ walk from this
village is a kind of spring from which the natives draw water; it
consists of two or three holes in excessively damp ground from
which water trickles and runs in insignificant streams to the
neighbouring heavily-wooded ground, which is thus rendered so
swampy that on entering, one is immediately immersed up to the
knees. There is thus no expanse of open water, but further on,
owing to the slope of the ground, the small streamlets unite to form
a brook—a secondary or tertiary tributary of the large river Luilu,
which itself is a tributary of the Lubilash-Sankuru.
I was much surprised to find numerous examples of G . palpalis
in this wooded swamp near the spring, as it was the first time that
I had met the fly under such conditions, namely, in a small wooded
swamp without the immediate presence of an uncovered mass of
water. Since then, however, I have often found G . palpalis even
in the smallest swamps, where the necessary vegetation existed.
Therefore, so long as a place possesses shade and a certain degree of
humidity, G. palpalis can exist, although it is not necessarily
present. Even permanent humidity is not essential, since temporary
humidity, as in the rainy season, will suffice. As soon as this
humidity disappears in the dry season, the fly disappears also, only
to reappear later when conditions are favourable. This reappearance
may be brought about in two distinct ways, firstly when conditions
become unfavourable in any region, G. palpalis is able to migrate to
a more favourable locality where there is permanent water, and later
to return to its former habitat with the change of the season;
secondly the conditions having become unfavourable to its existence
the fly dies, but there remain in the ground numerous pupae (for the
development of which dryness is required) which furnish adults for
the subsequent more suitable period. These two phenomena take
place, according to circumstances, separately or even simultaneously,
for the preservation of the species.
It is only necessary to recall what has been stated above, viz.,
that the district is bestrewn with marshes and swamps of various
sizes, that it is intersected with numerous watercourses, and that the
lower parts, where all these occur, are generally wooded, for us to
understand that the semi-ubiquity of G. palpalis in this region is
very natural. Happily, however, there are many exceptions to this,
520
although one may not always be able to explain the reasons for them.
In certain regions, for example in the territories of Kanda-Kanda and
Tshofa, G. palpalis occurs on the majority of the streams, while in
the Kasongo-Niembo region the fly is relatively rare. From an
examination of several rivers on numerous occasions and in different
places, I have been able to deduce the following rule:— If a known
spot on a river is positive as regards the presence of G. palpalis , then
any others situated doivn-stream , on the same river , are also positive
(provided always there is shade-giving vegetation); the character of
places situated up-stream is unknown , and may be positive or
negative. Further, if any place is negative, points higher up the
river are also negative, but points lower down are unknown.
DIFFICULTIES ENCOUNTERED IN EXAMINING A REGION FOR
QLOSSINA PALPALIS
The information here given on this subject may perhaps be of
some value to other medical men, and to those in general who are
interested in this question; it may even help them to avoid certain
errors.
On the 4th May, 1913, when travelling from Mwana-Tonto to
Tutu (via the Kanda-Kanda to Mutombo-Mukulu route) it was
necessary to cross the large river Luilu, which I reached at 7 o’clock
in the morning, one hour after sunrise. Rain had fallen on the
previous evening, and it was damp and fresh; no insects were
visible. About 7.30 the animal world commenced to awake; first
came the * ordinary’ flies, then some butterflies, afterwards various
hymenoptera (small bees, wasps, carpenter bees), and horse flies
(Tabanus ), but it was only at 9 o’clock that the first specimen of
G. palpalis appeared and not until 10 o’clock did they become
numerous. I would, therefore, not have seen a tsetse-fly if I had left
the river before 9 a.m., even had I remained there for a considerable
period, and consequently one might easily have stated that
G. palpalis was not present in this spot.
The same thing occurred on the River Lomami along the
Mutombo-Mukulu to Kasongo-Niembo route, where I arrived at
8 o’clock in the morning on the 24th May. It was the beginning of
the dry season, and was cold in the night and early morning; at this
521
hour (8 a.m.) the dew had not yet disappeared, it was very fresh
beneath the trees on the river banks, and all was quiet. I waited—
first appeared some ants on the tree trunks and some of the large
Tipulids, then successively * common* flies, small moths, butterflies,
carpenter bees and, at last, at 9 o’clock the first G. palpalis. It was
not until nearly 9.15 a.m. that the second appeared, but at 10 o’clock
these flies were numerous and commenced to bite.
I reached the River Lovoi (when travelling on the Kikondja-
Kabongo route) late in the evening of the 4th February, and installed
myself near the bank. It rained continually throughout the night,
and although, from an early hour in the morning, I repeatedly
visited the river bank, I had not seen a solitary tsetse-fly by
11 o’clock. Naturally under the circumstances, I should have
concluded that no G. palpalis were present here at all, but for the
fact that I remained in this place for the rest of the day and then
saw the flies appear in the afternoon. After heavy and prolonged
rain, then, during which the temperature is lowered and everything
saturated, G. palpalis remains hidden and inactive for a considerable
period.
The River Lukula is a tributary of the River Lubefu. It is
narrow and deep (about 5 metres in width) with a very rapid
current, and it flows between high and steep banks covered with
dense vegetation. The upper branches of the trees on each side
are in contact, and form a complete roof over the stream. I crossed
the Lukula, in its upper regions between Mwana-Kialo and Bukile,
on the 10th July, but was not able to discover a single G. palpalis
in spite of the fact that I remained in this spot for one hour at a
very suitable time of the day (11 o’clock to noon). On the
22nd July, I re-crossed the Lukula between Dibwe and Lubefu, that
is to say, not very far from the mouth. Every July a dense mist
occurs in the mornings, which only disperses later in the day, about
8 or 9 or even 10 o’clock. It was on such a morning that I reached
the river. In spite of the relatively late hour, 9 o’clock, the mist
had not yet cleared on the well-wooded banks of the river, and it
was still cool under the canopy formed by the upper branches of
the trees. I remained an hour on the river, but saw no traces of the
fly. In spite of this, and of the fact that I had not found
G. palpalis in the upper regions of the river under exactly similar
522
conditions, I was not satisfied. This spot was so near the mouth
that it would certainly be strange if G. palpalis were not present.
It was already between 9 and 10 o’clock, and elsewhere was quite
warm, but on the river it was still fresh since the sun’s rays had not
yet thoroughly penetrated the overhanging branches. Although
it is generally stated that this fly is active from sunrise, I waited,
and was well rewarded for doing so, as about 11 o’clock I heard the
typical and peculiar humming of the first tsetse-fly. I may mention,
in this connection, that when travelling on the River Lomami in a
native boat, between Tshofa and Gandu, from the 9th to nth
August, G. palpalis appeared about 7.30 a.m., in spite of the
morning mist. The Lomami, however, at this spot is more than
100 metres wide, and, consequently, its surface is quickly warmed
by the sun, and the fly appears earlier than on the Lukula.
At 7 o’clock on the morning of the 21st May, I crossed the large
river Luembe on the Mutombo-Mukulu to Kabongo route. No
specimens of G. palpalis were to be seen, and, unfortunately, I was
unable to remain for any length of time. Would it be more correct
to say in consequence that G. palpalis did not occur here, or that
it was not visible owing to the earliness of the hour ? The answer
is very simple. The previous evening, I stopped in a village
situated about half an hour’s walk from the river, and, foreseeing
what might happen, went in the afternoon to examine the river, and
there saw numerous specimens of G. palpalis.
Between the station Kanda-Kanda and the mission of Thielen
St. Jacques are three small streams, on all of which G. palpalis
occur! Between Thielen and the hamlet Pokote are also three small
streams and a little brook, and on all of these again the fly is
found. My two visits to these spots were paid between 10 a.m. and
1 p.m., and 10 a.m. and noon, respectively. When some days
later, in May, I crossed two small rivers, surrounded by forest
vegetation, between Mwana-Msenge and Mwana-Tonto no trace of
G. palpalis was to be found. The only difference was that I had
halted here early, and had crossed the streams between 7 and 8 a.m.
I could continue to give similar examples, but consider it
unnecessary, as the few facts cited prove sufficiently that much care
is required. It is evident that, if one remains for a long time in one
spot, and examines it on numerous occasions with the result that,
523
under all conditions, morning, noon and evening, in the dry season
and in the rainy season, no sign of the tsetse-fly occurs, then it can
be stated that the fly is absent. But without such precautions one
must not be too dogmatic. From a casual examination it is only
possible to say that G . palpalis has not been seen, but this does not
necessarily mean that it is not present. In fact the statement, that
this tsetse-fly is active from sunrise to sunset, is incorrect; G. palpalis
flies and bites during the day when the temperature is not too low
and when the surrounding vegetation is not too heavily saturated by
rain or dew. Other conditions being equal, G. palpalis appears
earlier and disappears later in the rainy season than in the dry
season, as the nights of the latter are relatively cold and the
evenings and mornings fresh. A little fine warm rain scarcely
affects G. palpalis at all, but during, and for a long time after,
heavy rain it is not in evidence. A severe night rain consequently
retards the appearance of the fly, and heavy rain later in the day,
or towards noon, causes it to disappear for the remainder of the day.
It is thus necessary to choose for the examination a suitable spot and
moment. During the rainy season there are mornings, afternoons,
and even entire days which are not favourable. Early morning and
evening are never suitable, especially in the dry season, and yet,
in the Congo, it is impossible to travel systematically at a period of
the day other than the early morning. In general, the presence of
G. palpalis is discovered quickly enough given a favourable time,
for example, mid-day, and often it is not even necessary to search
for the flies, as they soon make their presence known—especially
when they occur in large numbers. There are cases, however, when
one can only discover the tsetse-fly after searching for an hour or
more, as the following examples show.
The River Lukashi is an important tributary on the left bank
of the River Lomami (another tributary on the right bears the same
name), and I have seen G. palpalis in March, at the crossing of this
river on the Kisengwa-Kabinda route, below the mouth of the
River Loamba. In June, on going from Kabongo to Kabinda,
I returned to this river at a point higher up stream near the village
of Gongo. Here the river is 6 to io metres in width instead of
being 50 to 60 metres wide as on the route Kisengwa-Kabinda.
Near Gongo, moreover, it has high, steep wooded banks. I reached
5 H
the river at 11 a.m. when the sun was very powerful and the heat
great, and waited in vain for an hour among the trees on the bank
for the appearance of G. palpalis; it was not until a further half-hour
had passed that I heard the characteristic humming noise produced
by these flies when on the wing.
The River Lurimbi enters the Lomami at Tshofa and rises
near Kabinda, at the foot of the hill where the village of the
well-known chief Lupungu is situated. The route Kabinda-Tshofa
follows, more or less, the course of Lurimbi. When going from
Tshofa to Kabinda opportunities occurred of visiting several places
on this river, and on examination G. palpalis was found to be
present everywhere. I then decided to seek the fly in Kabinda
itself, and it was only after waiting a whole afternoon that I
succeeded in capturing two examples at the crossing of the river
situated about twenty minutes from the station.
It is, therefore, impossible to draw up an exact and detailed map
of the distribution of G. palpalis in this district, even along those
routes taken by me, in spite of the care, attention and interest that
I have taken in this question, for the fact of its not being found does
not, as we have seen, imply the absence of G. palpalis.
From the last two examples cited it is evident that much trouble
was involved in discovering G. palpalis on the passages of the
Lukashi and the Lurimbi. Both these routes—Kabongo-Kabinda
and Kabinda-Pania—are much traversed, and, in these cases
therefore, the view of Roubaud and other experts that G. palpalis
is especially abundant at river-crossings much frequented by animals
and men, apparently does not hold. I do not contest the accuracy
of the cases described by Roubaud and others, yet, although I have
seen G. palpalis at the river crossings on numerous occasions,
I have never observed that it was more abundant in such places
than in those which the natives seldom visit. It seems to me,
therefore, that strictly speaking these specialised haunts are far from
being demonstrated.
The question as to where G. palpalis rests during the night, the
colder mornings and wet weather is of interest. These hiding places
are evidently among the plants or under the leaves of trees, but
the exact positions I have been unable to discover. On several
525
occasions I have examined the under sides of the leaves of various
plants, but always without success.
All observers have verified, and many have given explanations of,
the fact that among series of captured specimens of G. palpalis the
males are far more numerous than the females. I have examined,
from this point of view, several of my collections of these flies from
various localities with the following results : —
No.
Date
Locality
Males
Females
Total
i
July
R. Sankuru (Pania) .
5 i
42
93
2
August
R. Lurimbi (Tshofa)
74
54
128
3
”
R. Lomami „ .
243
2°5
4+8
4
”
R. Buluy (Piani Tshungu) .
34
22
56
5
>>
In pirogue on the R. Lomami (Tshofa-
Gandu) .
83
42
125
6
May
R. Lubishi .
3 6
66
102
7
R. Lucmbe .
>4
1
15
8
March
R. Lukashi .
3»
11
42
1
9 ’
>>
R. Kela (Inga) route Kabinda-Kiscngwa 1
46
'7
63
IO 1
July
R. Lukula .
*5
25
i 4°
1
My own results thus, on the whole, support these statements,
but in two cases, Nos. 6 and io, the opposite occurs, viz., the
females, many of which were gravid, are almost twice as numerous
as the males.
I have noticed further, at various times, certain morphological
peculiarities among those specimens which I have collected. In
some regions they have been uniformly and remarkably small.
Dampness darkens the colour very greatly, and a moist specimen,
when dying or after death, becomes very dark and the abdomen
almost black, so that the segments can scarcely be discerned.
When the abdomen of the fly is distended, either by the larva
or by ingested blood, the dorsum of the abdomen becomes distinctly
526
paler and, owing to the distension of the intersegmental membrane,
appears regularly banded. Females are generally much larger than
the males, and it is in the former that the distension of the abdomen
with blood is most marked, as they are more voracious and are able
to ingest more blood at a single meal than the males.
Kabinda,
N ovember, 1913.
5 2 7
A NOTE ON A TRYPANOSOME OF THE
BLACK RAT (EPIMYS RATTUS)
BY
J. W. SCOTT MACFIE, D.Sc., M.B.
WIST AFRICAN MEDICAL STAFF
(Received j or publication 6 October , 1915)
Plate XXXVIII
The trypanosome, of which a description follows, was found in
the blood of a young black rat (Efitnys rattus ) sent to the
laboratory, Accra, on August 5th, 1915, by the Medical Officer of
Health, Dr. J. B. Alexander, to whom I take this opportunity of
tendering my thanks.
On examining the blood of this animal immense numbers of
trypanosomes were found to be present. The parasites were
evidently of the T. lewisi type, but even at first glance it was obvious
that they were extraordinarily polymorphic, and that some of
the forms were very large, and characterised by a remarkable
prolongation of the posterior end into a whip-like extension. The
trypanosomes were very active, and when progressing moved rapidly
across the field of the microscope. Their movements were of two
kinds. The one kind, a large wriggling movement involving the
whole body, was that seen when the parasites were progressing; the
other kind was exhibited when the trypanosomes were stationary,
and consisted in a very rapid vibration of the anterior end. By
means of the latter movement the blood corpuscles were set rotating
on their own axes, and at the same time revolving in a continuous
stream round and round the anterior end of the trypanosome. The
movement was such as might have been produced by the flagellum
being wrapped round the corpuscles, and then suddenly withdrawn,
much as the string is drawn from a whip-top; but owing to the
activity of the movements it was impossible to make out how they
were actually brought about.
In stained specimens the polymorphism of the parasite was seen
5 z8
to be extreme. Some of the trypanosomes were very small, others
very long; some slender, others broad. A hundred individuals,
taken as they came, were drawn with the camera lucida, and
measured by the tangent line method. The longest of these
measured 48//, the shortest 15 //, and the average length was 30*02/1
(see Tables 1 and 2). The majority, fifty-five, were between 30/t
and 39 fi in length. In breadth the trypanosomes averaged 2*62/1,
and ranged from 2/1 to 6/1; but the great majority were about 2 fi
broad (see Table 3).
From such a small number of measurements it is of course
impossible to plot a reliable curve representing the distribution of the
trypanosome according to length. Neither are the extremes repre¬
sented in these few measurements. Elsewhere in the films parasites
were measured which reached a maximum of 52/1 in length on the
one hand, and on the other a minimum of 12fi. Some aggregates of
division forms were also seen, similar to those found, in T. lewisi
(Plate XXXVIII, fig. 8).
Certain features were common to all the forms of the trypanosome.
The body was in every case prolonged posteriorly for a considerable
distance beyond the micronucleus; the cytoplasm was seldom or
never granular; the micronucleus was large and oval or rod shaped;
the nucleus was rounded or oval, and usually situated well in the
anterior part of the body; the undulating membrane was but slightly
developed; and there was always a well marked free portion to the
flagellum. Four types of parasite could be distinguished, but
between them, and linking them together, every intermediate stage
could be found; and for this reason I believe that the rat was infected
with only a single species of trypanosome. The majority of the
trypanosomes were about 30/1 to 35/1 in length, and 2/1 in breadth
(figs. 7, 10). The cytoplasm stained a reddish-blue colour by
Leishman’s method, and was without definite granules. At the
anterior end there was a well marked free flagellum, and the posterior
extremity was prolonged for a considerable distance beyond the
micronucleus. Very much smaller trypanosomes, some of which
measured only 12 fi in length, which stained similarly, were also
fairly common (figs. 18, 16). In them the posterior end tapered
rapidly into a finely pointed cone, and the micronucleus was not
infrequently situated alongside of, or even slightly anterior to, the
529
Table I.—Distribution, by percentages, in respect of length of a trypanosome of the black rat.
Length in microns
*5
i
16 j 17
18
19
20
21
22
23
!
2 4 25
26
27
28
29
3 o
3 *
i
I 0
1
3
i
4
5
0
O
3 ;
i
!
3 i 2
1
4
3
. 3
8
7
8
Length in microns
32
33
34
35
36
37
38
39
40 41
42
43
1 ;
44 1 45 1 46
I 1
47
48
6
10
6
|
,0
5
1
1
1
1
° } °
1
1
2
1 i
0 j 1 0
1
O
i
I
Table II.—The lengths of 100 trypanosomes from a black
rat distributed in groups of ten microns each.
Length in microns
10-19
20-29
30-39
40-49
9
3»
55
5
Table III.—Breadths, to the nearest whole numbers, of
ioo trypanosomes from a black rat at Accra.
Breadth in microns
<>3
20
4
2
macronucleus (fig. 15). Larger forms were also common. They
were broad trypanosomes, sometimes measuring 6 fi at their widest
part, with cytoplasm that stained a bright blue, and showed an
alveolar structure (figs. 4-6). The posterior end was prolonged
beyond the level of the micronucleus as a broad-based cone, and the
anterior end terminated in a well developed free flagellum. The
macronucleus was placed anteriorly, the micronucleus a little to the
posterior side of the middle point. There was generally a
conspicuous vacuole just anterior to the micronucleus. These three
types were very similar to those figured by Delanoe (1915) as typical
of T. eburneense; the first type representing the adult, and the other
two the multiplicative forms. In addition, however, in this rat
examined at Accra, there were some very remarkable forms with the
posterior end prolonged into a whip-like extension (figs. 1-3). These
trypanosomes stained a reddish-blue colour, and measured up to
52/1 in length, and 3 n or 4 fi in breadth. Intermediate forms were
found which seemed to link this type with the commonest form of
trypanosome present in the blood. The anterior end terminated in
rather a short free flagellum; the macronucleus was situated in the
anterior part of the body, and the micronucleus, which was rod
shaped, lay right across the posterior end at the point where the
whip-like extension might be considered to begin. Beyond the
micronucleus the posterior end of the body was drawn out into a fine
flagellar filament measuring in some instances 24/1 in length. The
following are the detailed measurements of three individuals of this
type: —
Free portion of the flagellum .
6 fi
6m
5 h
Anterior extremity of the body to the middle
of the macronucleus.
8 M
IO fJL
IO n
Middle of the macronucleus to the micronucleus
12 M
IO n
12 n
Micronucleus to the posterior extremity of the
body .
21 f 1
24 M
j 2 3 h
Total length.
47 M
3° M
50 M
Breadth at the widest point .
3’5 M
4 M
2-5 p.
Such forms are similar to those found by Lingard (1906) in
Mus niveiventer and M. decumanus> for which the name T. Ion go-
53 *
caudense was originally proposed, but which subsequent observations
have proved to be forms of T . lewisi which are ‘ of constant
occurrence and very numerous at a certain stage of the multiplication-
period 1 (Minchin, 1912).
On August 6th, the day after the rat was first examined, the
forms of trypanosome found in the peripheral blood were similar to
those described above; but on the following day the T. longocaudense
forms and the very small forms were exceedingly rare. On
subsequent days only the first type, trypanosomes of the adult form,
was represented. At this period of the infection the trypanosomes
were on the average somewhat longer than typical examples of
T . lewisi , but the difference was of a degree that could scarcely have
been appreciated without measurements. The average length of
twenty-five individuals that were drawn and measured by the
tangent method was 33 fi.
Delanoe (1915) has recently published an interesting account of
the trypanosomes found by him in the course of the examination of
600 rodents at Bouake, on the Ivory Coast. One of the species
described, that for which the name T. eburneense is proposed,
appears to resemble closely the trypanosome described above, with
the exception that none of the large forms with the whip-like
extension of the posterior end seem to have been present. It is
hardly possible that such remarkable forms could have been over¬
looked, but it is possible that in the animals examined the infection
had advanced beyond the stage at which these forms are present in
the blood; for T. eburneense is a trypanosome of the T . lewisi type,
and showed other multiplicative forms similar to those of T. lewisi ,
and may therefore in all probability be assumed to show at times the
forms with the posterior end greatly extended.
Thirty-six adult trypanosomes from naturally infected Mus concha
were measured by Delanoe. The average length was 34*9^, and the
range from 32 n to 38 fi. These measurements are similar to those
of the commonest form of trypanosome found in the black rat
examined at Accra, which were from 30 fi to 39 fi in length, and
which constituted 55 per cent, of the forms present on the first day
on which the rat was examined. It would appear probable therefore
that the trypanosome found in the black rat at Accra was of the same
species as that found in Mus concha by Delanoe, and described by
him under the name T . eburneense.
532
The question then arises is this trypanosome sufficiently distinct
to be entitled to specific rank or should it be considered as merely a
variety of T. lewisi. Delanoe concluded that his measurements
showed that the trypanosome of Mus concha was decidedly longer
than T. lewisi. It may be doubted whether the measurement of
only thirty-six trypanosomes at an unknown stage of the infection is
sufficient evidence for such an assumption. T. lewisi measures,
according to Laveran and Mesnil, 24 fi to 25 n in length, but other
observers have given somewhat higher figures, and three individuals
measured by Delanoe himself were found to be 32/4, 32/1 and 29 fi
long respectively. Other trypanosomes are known to show marked
variation in average length at different stages of the infection, and
also when introduced into different hosts; and there would appear
to be no reason why T. lewisi should be an exception to this rule.
T. eburneense was found in six specimens of Mus concha , all of
which were somewhat heavily infected; and by means of inoculations
the parasite was transmitted to Golunda campanae , * rats savanes/
and Xerus erythropus , but striped rats (rats rayes), white rats, and
guinea-pigs appeared to be refractory in the few experiments that
were made. The trypanosome appeared to possess some degree of
pathogenicity, especially when inoculated into Golunda campanae .
The trypanosome found at Accra was inoculated into only one
white rat, and one guinea-pig, but no infections were thus
transmitted. It is difficult to decide what degree of importance
should be attached to such inoculation experiments since T. lewisi
has been transmitted to various animals besides rats, and it does not
appear to be known whether Mus concha , and the other animals to
which Delanoe transmitted T. eburneense , are susceptible, or what
effect if any is produced on the morphology of this trypanosome
when introduced into such unusual hosts. It is interesting to note,
however, that Brown (1914) concluded, as a result of his work on a
pathogenic strain of T. lewisi , that ‘ morphological anomalies were
most pronounced in infections that showed unusual conditions of
multiplication and that such infections usually proved severe/ an
observation that may perhaps be interpreted as supporting the view
that trypanosomes of the T. eburneense type may be varieties of
T. lewisi.
533
REFERENCES
Brown, Wade H. (1914)* Morphological and Developmental Anomalies of a Pathogenic
Strain of T rypattosoma lere is i and their Relation to its Virulence. Jl. Experim.
Med ., Vol. XIX, p. 562.
Delanoe, P. (1915). Au sujet dcs trypanosomes du type T. lervisi Kent rencontres chez
des Muridcs dans la region de Bouake (Cote d’Ivoire). Bull . Soc. Path. Exot. f
Vol. VIII, pp. 80-88.
Laveran, A. et Mesnil, F. (1912). Trypanosomes et Trypanosomiases. Paris. Masson
et Cie.
Lingard, (1906). Jour . of Trop. Vet. 6V1., Vol. I, p. 5. (Quoted by Laveran and Mesnil).
Minchin, E. A. (1912). Study of the Protozoa. London, Edward Arnold.
53 +
EXPLANATION OF PLATE XXXVIII
Figs. 1-18. Trypanosomes from a black rat (Epimys rattus )
examined at Accra, Gold Coast, West Africa, x 2000.
Annuls Trop. Med. & Parasitology, Vol. IX.
PLATE XXXVIII.
A TRYPANOSOME of EPIMYS RATTUS.
S 3 S
THE ARTIFICIAL CULTIVATION OF
HANSEN’S ‘BACILLUS’
BY
H. BAYON, M.D.
(Received for publication 9 October , 1915)
It is quite evident that a conclusive answer to the objections and
criticisms of Fraser and Fletcher (see these Annals , July, 1915,
p. 381) could only be given by discovering a method of isolating
and artificially cultivating Hansen’s ‘bacillus/ not only with its
original morphology, but also in a relatively easy fashion. No such
technical refinement is known at the present moment.
A long array of negative results is no doubt a formidable
argument in the hands of competent bacteriologists, but both human
and rat leprosy have shown similar anomalies in relation to
problems which were apparently much easier to solve than the
isolation, in pure culture, of a micro-organism which belongs to a
group of bacteria known to resist artificial cultivation in a most
persistent fashion.
Though the communicability of leprosy has been established as
a result of extensive epidemiological observations, we know that
hundreds of ward attendants have worked, often for years, in
asylums where hygienic precautions were practically absent and yet
did not contract the disease. Therefore, following the line of
argument adopted by some critics, contagion does not exist.
The long series of sterile culture tubes found by Fraser and
Fletcher are not without precedent. Similar numerous attempts
were carried out by K. F. Meyer before he succeeded in isolating
the acid-fast micro-organism of the cattle disease which he calls
enteritis hypertrophica bovis specifica, and which presents several
features in common with leprosy.
536
If all the diphtheroids isolated from leprous nodules are
contaminators, as implied by the positive assertion of Fraser and
Fletcher, then surely all must show the well-known characteristics of
the common Corynebacteria found on the body-surface. If, as many
other observers contend, they are, at least in some instances, related
to Hansen’s 1 bacillus/ then they must be distinguishable by means
of sugar-tests and cultural appearances from any other similar germ.
I take it, therefore that, though they do not mention the fact,
Fraser and Fletcher have assured themselves, by means of the usual
bacteriological methods, that the diphtheroids they isolated were
identical with some well-known germ cultivated from the skin.
However, I have already acknowledged that the destructive
criticism of Fraser and Fletcher can only be met by piling up
experimental evidence and arraying further observations and results.
At the present moment I am precluded from attempting anything in
this line, but I may be allowed to point out that my last paper in
these Annals affords an answer to one of the questions set up by my
critics in a paper they published in the Lancet recently.
Dealing with Kedrowsky’s culture, Fraser and Fletcher (1915)
ask: 'Why, if this is a culture of the leprosy bacillus and it can
produce leprosy in animals, did neither investigator produce similar
lesions by the inoculation of emulsions of leprous tissue ? Bayon,
at least, had abundance of material. We have performed such
experiments with uniformly negative results.’
My answer is that at Robben Island, though I had abundance
of material at my disposal, I only succeeded in one single instance
in producing lesions in a rabbit which lasted any length of time,
in this case one year and seven months. All other experiments on
rats, though I inoculated over a hundred, did not succeed to my
satisfaction. In London, where I only had three cases of leprosy at
my disposal, I succeeded (see Plate III, fig. 14, in these Annals ,
Vol. IX). The lesions produced are absolute counterparts of the
deposits brought about in some cases by the injection of Kedrowsky’s
‘ bacillus.* But, as stated, whether one injects ground-up nodules or
Kedrowsky’s culture, in the great majority of cases the bacteria get
simply eliminated, without leaving any visible trace. In single rare
instances, they produce bacillary deposits similar to those found in
the inner organs of some lepers.
537
I am compelled to admit that Fraser and Fletcher are equally
uncompromising when interpreting the results of their own experi¬
ments. On page 15 of the Lancet of July 3rd, 1915, they report on
some interesting and promising injections of emulsified leprous
tissue into guinea-pigs, which deserved being followed up by a
sufficiently long observation of the animals, in addition to an
attempt to transmit the infection to second or third generations.
But the fact that not a trace of a nodule could be found anywhere
after three weeks and that the organs were apparently normal, leads
these investigators to the conclusion that the deposits of acid-fast
rods they found in the spleen, liver, and in a peritoneal gland did
not carry any conviction.
I may be allowed to say that, after having done or attended at
over ninety autopsies of lepers at Robben Island, in only one single
case did we find on microscopic examination acid-fast bacillary
deposits in the liver resembling in quantity, intracellular and
extracellular situation the well-known lesions of the skin leproma.
We cannot expect skin lesions in animals inoculated with leprosy:
all we can hope for are discrete deposits in the inner organs. If
they can be transmitted through some generations and persist for a
considerable time, and the bacillary deposit is superior to the
quantity injected, then it seems to me that by all the laws of
experimental medicine the inoculation has succeeded. This is the
case with one experiment fully described in my paper.
It cannot too often be repeated that the scanty positive results
obtained in the experimental study of leprosy are absolutely in
keeping with what we know of the clinical features of the disease,
its low and eminently capricious infectivity; but that here, more than
when dealing with any other disease, the partial and incomplete
interpretation of hundreds of negative observations cannot invalidate
the proof positive of a single successful inoculation.
538
REFERENCES
Bayom, H. (1915). Leprosy: a perspective of the results of experimental study of the disease.
Annals Trop. Med. & Parasitol.. IX, pp. 1-90. Six plates.
Fraser, H., and Fletcher, W. (1913). The Bacillus leprae: has it been cultivated ? Lancet ,
Sept. 27, 1913, pp. 918-921.
- (1915). The cultivation of the Leprosy Bacillus. Annals Trop. Med. & Parasitol.,
IX, pp. 381-382.
- ( X 9 1 S)« Leprosy and Kedrowsky’s Bacillus. Lancet , July 3, 1915, pp. 13-16.
539
STUDIES IN BLACKWATER FEVER*
V.—THE DURATION OF HAEMOGLOBINURIA
BY
J. W. W. STEPHENS, M.D. Cantab.
SIR ALFRED JONES PROFESSOR OF TROPICAL MEDICINE, THE UNIVERSITY OF LIVERPOOL
(Received for publication 28 October , 1915)
With Chart
I have collected the data on this point in 167 cases. They
include cases in which a relapse had occurred, the relapse being
considered as a fresh attack. I have in previous studies drawn
attention to the difficulty of obtaining exact records of various
symptoms, and in the following analysis have only been able to
classify the figures in somewhat wide intervals. As they stand,
they are I think of interest, and if they are confirmed by further
observations will at least tend to make our conception of the
blackwater process more precise. It is possible too, that a study
of the data with regard to the duration of haemoglobinuria together
with a study of the temperature curves might reveal points of
importance, and further a comparison with the data, if such be
available, for the paroxysmal haemoglobinurias of adults and
children and for the parasitic haemoglobinurias of animals might
reveal points of difference or agreement which would throw
light on the nature of the factors involved in the production of
haemoglobinuria. At present I simply record the facts so far as
I have been able to ascertain them.
• Part I : Annals of Trop. Med. & Parasite 1913. Part II : Jhid 1914. Part III : Ibid,, 1915.
Part IV : Ibid., 1915.
540
Table I.—Duration of Hacmoglobinuria.
Hours
Hours
Hours
Hours
Hours
Hours
Hours
Hours
Total
Duration .
—
1-4
4-8
8-12
12-18
18-24
* 4 - 3 6
36-48
—
Cases .
3 *
11
9
22
9
26
22
20
122
Days
Days
Days
Days
Days
Days
Days
Days
Duration . 1
—
2-3
3-4
4-5
5-6
7
8
—
—
Cases .
—
3 °
7
5
I
1
■
45
1
167
* Haemoglobinous urine passed once only.
We may arrange these data in the following way : —
Table II.—Duration of Hacmoglobinuria.
Duration
Cases
Per cent.
1 day or less .
80
47*9
1-2 days .
1
42
25.1
2-3 days .
3 °
17-9
3-4 days .
7
|
4 *i
4-5 da y* .
5
2-9
5-6 days .
|
>
—
7 d »y» .
1
—
8 days .
1
—
167
—
In the 80 cases in which the hacmoglobinuria lasted i day or
less, the duration was 0-12 hours in 45 cases, 12-24 hours in 35.
5+2
We may also group the data into the following periods: —
Table III.—Duration of Haemogtobinuria.
Duration
Cases
Per cent.
Not more than 2 days ... .
122
73
More than 2 days .
45
26-94
Not more than 1 day .
80
47-9
More than one day .
87
5 *-i
Not more than 12 hours .
45
26-94
More than 12 hours .
122
7 )' 06
Approximately, therefore, in these 167 records,
The duration is not more than 12 hours in a quarter of the cases.
,, ,, ,, 1 day in half of the cases.
,, ,, ,, 2 days in three quarters of the
cases.
543
SOME EXPERIMENTAL RESEARCHES
ON INDUCED HERPETOMONIASIS IN
BIRDS
BY
H. B. FANTHAM, M.A. Cantab., D.Sc. Lond.
LECTURER ON PARASITOLOGY. LIVERPOOL SCHOOL OF TROPICAL MEDICINE
AND
ANNIE PORTER, D.Sc. Lond.
BEIT MEMORIAL RESEARCH FELLOW, QUICK LABORATORY, CAMBRIDGE
(Received for publication I November, 1915 ’
Plate XXXIX
CONTENTS
PAGE
I. Introduction. 543
II. Material and Methods . 544
III. Experimental Work. 545
IV. The Morphology of the Parasites in the Insect and tiie Avian Hosts ... 549
(a) Herpetomonas jaculum in the Ncpa and the Canary . 549
(b) Herpetomonas culicis in the Culex, Sparrow and Martin. 550
V'. Natural Herpetomoniasis in Birds . 552
VI. The Flagellate Stage in Herpetomoniasis and Leishmaniasis . 553
VII. General Conclusions . 555
VIII. Summary .. 556
References . 556
Explanation of Plate . §58
I. INTRODUCTION
The significance of certain insects in relation to disease has been
recognised for some time past. That plague is conveyed by fleas,
relapsing fever by lice, malaria by mosquitos, and sleeping sickness
by tsetse-flies, has become common knowledge. Many insects other
than the above serve as hosts for various protozoal parasites, but the
latter are usually regarded as innocuous to vertebrates, more
especially if the insect host is non-sanguivorous. There are,
5+4
however* a number q£. obscure diseases of which the excitants have
not been determined, and also a number of flagellates normally
parasitic in insects whose pathogenicity towards vertebrates has
never been put to the test.
In continuation of our work on the introduction of insect
flagellates into vertebrates, we have, for some time past, been testing
the pathogenicity of certain insect flagellates with respect to birds,
the last great group of the European vertebrates that has remained
untested by us. The present paper records the successful infection
of birds with two insect flagellates, Herpetomonas jaculum , Leger,
from the water scorpion, Nepa cinerea , and H. culicis, Novy,
MacNeal and Torrey, from Culex pipiens . The testing of the
pathogenicity of insect flagellates towards birds has been of interest
for two reasons. First, in May, 1907, Drs. Edm. and Et. Sergent
recorded briefly and figured a herpetomonad that they had
found occurring naturally in the blood of a pigeon in
Algeria. In the second place numbers of birds die annually from
other than old age and wounds, and the cause of death is never
ascertained. Some of our experiments, coupled with the fact that
the crops of the birds frequently contain insects, suggest that
undetected herpetomoniasis may be the cause.
We have referred in our previous papers to the work of Laveran
and Franchini on the introduction of insect flagellates into mammals.
Our own experiments were commenced early in 1911 with rats,
which were refractory to the introduced herpetomonads. However,
our experiments were continued, different hosts being used with
more successful results. We have much pleasure in thanking
Professor G. H. F. Nuttall, F.R.S., for his kind interest in our
researches, and for looking at many of our preparations.
II. MATERIAL AND METHODS
The birds used were canaries ( Seritius catiarius ), sparrows
(Passer domesticus)> and martins ( CJtelidon urbica). The canaries
were bought from a bird-dealer, the sparrows and martins were
caught, and were practically tame when used for feeding or
inoculation. The control birds in each case are still alive and
healthy, or were found healthy when killed.
S 45
The birds were fed either with the entire insects (containing
herpetomonads) or with their alimentary canals that had been
removed. There was usually no difficulty over the feeding. After
the infective feed had been given, grain and shredded cooked meat
or egg were given as food. Infection of birds by inoculation was
also tried.
The insects {Nepa cinerea and Culex pipiens ) were obtained
chiefly from the neighbourhood of Cambridge, and some of the
Culex larvae were bred out and identified. For the present we
retain the species names of the various herpetomonads (e.g. H. culicis
and H . jaculuni) as given originally, though we have much evidence
that such are not true species, but are rather physiological races of
one or two distinct species.
Blood smears of the experimental birds were taken at intervals.
Smear preparations of the organs were made at death, fixed wet
with osmic vapour followed by absolute alcohol, or with Bouin’s
fluid, and stained with Giemsa's solution or iron-haematoxylin.
The birds used were carefully examined for ectoparasites, but
none was found. Blood examinations for haematozoa at the
commencement of the experiments were equally negative.
III. EXPERIMENTAL WORK
Herpetomonas jaculum has been shown by us to be capable of
infecting certain fish, amphibia, snakes and mice. It was therefore
tested with birds. The use of H. culicis was suggested by finding
many Culex remains in the crops of some sparrows and martins
found dead and sent to us for examination. They had been too
long dead to allow of detection of blood parasites, had any been
present. In the light of our experiments it is possible that such
parasites may have been present.
Experiment i (H. B. F.). A canary, 9 , was fed with material
containing Herpetomonas jaculum. The bird snapped the dissected
intestines of two Nepa cinerea containing H. jaculum somewhat
greedily, and also swallowed two infected nymphs. It was then fed
on ordinary, mixed bird seed and chopped egg until the day of its
death, which occurred 51 days later. Blood smears were made at
intervals, during the period of the experiment. At first the bird
seemed quite healthy. Twenty-one days after the commencement
of the experiment, a blood smear showed the presence of elongating,
post-flagellate forms of H. jaculum. About a fortnight later the
bird became very mopy, sat huddled up, refused food and shivered.
Again parasites were found in the blood. The attack lasted three
days, when the bird seemed to recover. Nine days later, on the
50th day of the experiment, the bird became mopy again. On the
51st day it seemed better at first, refused its seeds, but ate the egg
supplied it. During the day it was brighter again, but in the
evening again refused food. It was hopping about half an hour
before its death, which took place the same evening quite suddenly
and with no struggle of any sort.
At the commencement of the experiment the bird weighed
26 grams; at its death it weighed io*2 grams.
At post-mortem, the body was found to be much emaciated.
The liver seemed normal, the spleen slightly enlarged. Only a very
small quantity of bone-marrow was present. The suprarenal bodies
were very hard and firm, much more so than in normal birds.
Smears were made of all the organs. When examined the results
were as follows:—Leishmania-like bodies and some uninucleate
forms were found in the liver, which also contained a few elongating
parasites and still fewer typical, flagellate herpetomonads. The
spleen smear showed a few uninucleate and leishmaniform
organisms, some of them intracellular. The heart and lungs
harboured leishmaniform bodies, and in the lungs developing forms
showing the root of the flagellum were seen. Smears of the kidney
showed the presence of a few uninucleate forms and some leishmania-
like parasites, and similar bodies were present in the bone marrow.
No parasites were found in the suprarenal bodies.
A control bird was killed, but no parasites were found in its
organs, nor had it lost weight.
EXPERIMENT 2 (A.P.). This experiment was undertaken as an
examination of the crop contents of several sparrows found dead
had shown the presence of a number of the common gnats,
Culex pipiens . These gnats are known to harbour Herpetomonas
culicis. A number of larvae and of adults were obtained, and were
fed to a 9 sparrow, which was fairly tame. The insects were taken
by the bird without any trouble. Four days later the bird became
mopy and had a shivering attack, which soon passed off. A fresh
547
blood film showed a single non-flagellate herpetomonad. On the 9th
day of the experiment the bird died suddenly. The liver and spleen
were softish, the kidneys, suprarenal bodies and lungs seemed normal.
The bone-marrow was small in quantity, but more fluid than usual.
The pancreas appeared to be enlarged. The ovary was large and
contained numerous ova. No protozoal parasites were found post¬
mortem in blood examined fresh or in the intestine. The bird
weighed 227 grams at death, showing a loss of 2*2 grams from the
commencement of the experiment, and exhibiting some emaciation.
At death, smears were made of the organs. The stained smears
showed that, as in the case of the canary, there was a generalised
infection of the parasite, in this case, Herpetomonas culicis . The
flagellate form of the herpetomonad predominated here. The non-
flagellate stages were less common. Transitional forms also were
present. The heart, liver, lungs, kidneys, and suprarenal bodies
contained well-developed flagellates. Leishmaniform bodies were
also present in smears of these organs, but were less frequent than
the flagellate forms. The bone-marrow also contained them as well
as elongating parasites. Leishmaniform elements in process of
division were found in the heart and liver, dividing elongating but
non-flagellate herpetomonads occurred in the bone-marrow, while
fully developed flagellates in various stages of division were present
in smears of the heart, liver and lungs.
Blood smears taken during the course of the experiment, and
stained, showed no parasites except on the day before death. A
smear taken 14 hours before death contained a single leishmaniform
element in process of elongation. The blood showed signs of
anaemia, and was very fluid.
The control sparrow is still alive and active.
Experiment 3 (H.B.F.). A young adult 3 martin was fed
with larvae and mature Culex pipiens containing Herpetomonas
culicis . It lived twelve days after the infective feed. The condition
of the bird at death and the distribution of the parasites was
practically the same as that of the sparrow, given in Experiment 2,
so that the description need not be repeated. Blood smears taken
during the course of the experiment were negative.
Experiment 4 (A.P.). A young adult $ martin was inoculated
subcutaneously with H. culicis . The bird was greatly frightened
548
and died after two days. No infection appeared to have taken
place.
EXPERIMENT 5 (A. P.). A young canary was fed with the
faeces of Nepa cinerea , the excrement, which contained H. jaculum ,
having been collected on a slide and mixed with bread. Blood
examinations were made at intervals. The bird lived 17 days after
the infective feed. A few non-flagellate forms were found on the 7th
and nth days. At post-mortem, a few non-flagellate forms were
found in the liver and spleen, elongating parasites in the liver and
bone marrow, and a few flagellates in the liver. The heart-blood
and tissue contained some multiplicative forms.
EXPERIMENT 6 (H. B. F.). A young mature martin, c?, was fed
with the faeces of several larvae and adults of Culex piptens y mixed
with small quantities of boiled mutton. The Culex faeces contained
post-flagellate or encysted stages of H. culicis. The bird lived 32
days after the infective feed. The body was somewhat emaciated at
death. Non-flagellate forms of H. culicis were present in the spleen
and lung, elongating forms in the bone-marrow and a very few
flagellate stages in the spleen.
EXPERIMENT 7 (H. B. F.). A 9 sparrow was fed with the faeces
of 'Nepa cinerea containing H. jaculum. On the nth day after the
infective feed, a probable parasite of the elongating flagellate type
was seen in the blood, but no others have been observed since. The
bird is growing somewhat thinner but is still alive at the time of
writing, and infection appears doubtful.
EXPERIMENT 8 (A. P.). A 9 canary was fed with food con¬
taminated with H. culicis. As usual, blood smears were taken at
intervals. No parasites were found. After 80 days, the bird was
killed, but no herpetomonads were found on examination of organ
smears.
In connection with these experiments, it should be remembered
that the flagellates of the insect hosts rarely co-existed with many
bacteria. In common with certain other workers, we found that when
the insects contained many bacteria, the protozoal flagellates usually
disappeared. Further, some digestion experiments performed by
us have shown that many bacteria introduced into the digestive
fluids of the bird’s stomach are destroyed by the same, while the
• flagellates are but little affected. This is not surprising, since
certain protozoal infections of man are known to flourish in an acid
medium, and are combated by the use of alkaline substances.
549
IV. THE MORPHOLOGY OF THE PARASITES IN THE
INSECT AND THE AVIAN HOSTS
The morphology of Herpetomonas jaculum and H. culicis in the
respective insects and birds is of interest. Little morphological
difference can be found in either case. It may be noted that when
the infection was of the chronic type, as in the canary (Expt. i), the
non-flagellate, leishmaniform bodies preponderated in the smears of
the organs, while the mature flagellates were more numerous in the
cases of the sparrow and martin (Expts. 2, 3), where the herpeto-
moniasis was of the acute type. We would point out that while such
was the case in these experiments of ours, we do not consider that
any generalisation can yet be made therefrom. However, it may be
noted that Monge (1914), dealing with flagellate stages of
Leishmania tropica in man in Peru, states that the presence of such
flagellate stages may be an indication of increased virulence. More
experiments are needed, and some are now in progress.
A. Herpetomonas jaculum IN THE NEPA AND THE CANARY
Herpetomonas jaculum parasitic in Nepa cinerea shows much
variation in size. The non-flagellate stages are oval, and show a
nucleus and well marked blepharoplast. The position of the
blepharoplast varies. These non-flagellate forms elongate, the
extension often being preceded by division of the nucleus and
blepharoplast. The root of the flagellum differentiates and finally
reaches the exterior, forming the free flagellum. The body
also elongates, and the mature flagellate is thus produced.
When change of host is necessary, retrogression and absorption of
the flagellum occurs, the body concentrates, and a cyst wall, at first
gelatinous but later shrinking to a thin, vamish-like coat, is
produced. This is the post-flagellate or encysted form of the
parasite, adapted for life outside the body of the host. When
ingested by a fresh host, it becomes the leishmaniform, pre¬
flagellate organism with which the cycle commenced.
Herpetomonas jaculum , as found in the experimentally infected
canary (Expt. 1), showed non-flagellate and flagellate forms, in
various stages of growth and division. Non-flagellate forms (Plate
XXXIX, figs. 1-15) were often from 4/x to 6*6// long by 2// to 5//
broad. Usually they were found singly (figs. 2-10), sometimes
550
clusters of two (fig. 11) were present, and on one occasion only a
number of somewhat narrow forms (fig. 12) was found. The body
cytoplasm was slightly alveolar. The nucleus usually was finely
granular and homogeneous (figs. 3-6); more rarely a karyosome was
seen (figs. 2, 7). The blepharoplast occupied different positions in
the body (figs. 1-10). It was often barlike (figs. 14, 15), sometimes
rounded (figs. 7, 17). No differentiation of structure was seen
within it. Dividing non-flagellate parasites were not numerous.
The elongating herpetomonads presented much the same structure.
In some of them the root of the flagellum was seen (figs. 16, 17).
This structure, as mentioned by one of us in a previous paper, was
first described by Mesnil and colleagues (1904) for Leishmania
tropica , by Christophers (1904) for L. donovani and by Novy (1909)
for L. infantum.
Full-grown flagellates (figs. 18, 19) were relatively rare in the
canary at the time of its death. It is possible that they may have
been more numerous at some period of its life, but that could not
be ascertained, the organisms usually occurring in the internal
organs of the host. Morphologically, the flagellates were like those
in the insect host, but the maximum size, as in previous experiments,
was not attained, though a flagellate was found with a body-length
of 20/1 .
From the foregoing it is seen that there is a close resemblance
between H . jaculum in the insect host and in the avian host into
which it was introduced by feeding, while the non-flagellate forms
recall those of the various leishmaniases of man. Further statements
regarding this subject will be made later.
B. Hcrpetomonas culicis IN THE CULEX, SPARROW AND
Martin
The life-cycle of Herpetomonas culicis in Culcx spp. resembles
that of H . jaculum morphologically, and need not be recapitulated
here. As found in the avian hosts, the non-flagellate forms of
H. culicis were oval or pyriform bodies (figs. 20-25), usually
measuring 4 /i to 6// by 2 /i to \/i. Occasionally, rarer, larger forms
were encountered (fig. 26), but it is possible that they were about
to divide. Non-flagellatcs in various stages of division were found
(figs. 27, 28).
55i
The flagellates were elongate (figs. 30-38), with both ends more
or less rounded, though the posterior end was often the more pointed.
The body length usually varied from 11 // to 16/4, and the breadth
from 1*5 n to 3'6 /k. The flagellum was often as long or longer than
the body. Still longer parasites (fig. 38) might be found. Some¬
times forms about to divide were wider (fig. 29), as would be
expected.
Multiplication of non-flagellate forms by binary fission was
found in preparations of the heart and bone-marrow, while dividing
flagellates (figs. 39, 40) were present in the smears of the heart and
lung. Rosettes of parasites, produced by repeated binary fission
in the insect host, were not found in the bird smears.
The nucleus of H. culicis was round (figs. 22, 33) or oval
(figs. 25, 31, 36), and was most frequently granular (figs. 21-32).
Occasionally a nucleus showing a karyosome was encountered
(fig. 33). The blepharoplast was always conspicuous, and varied
from round (figs. 31, 36-38) to bar-like (figs. 34, 35). Meta-
chromatic granules (fig. 35) were not common in the cytoplasm,
which was alveolar. Traces of contractile myonemes (fig. 33) were
occasionally seen.
The morphology of the parasite in the vertebrate host, then, is
practically identical with that of the Protozoon in the gnat. The
plasticity of the organism is apparent, and the capacity for morpho¬
logical variation in the parasite coincides with the increase in its
pathogenicity to its new hosts.
The results of the foregoing experimental work show that
Herpetomonas jaculum and H. culicis can parasitise and be
pathogenic to birds to which they gain access. The adaptability
of the flagellates to life in vertebrate hosts is well marked. They
exhibit the same morphology as when in the insect host, and attain
almost the same dimensions. The degree of pathogenicity induced
appears to vary, the disease resulting being of a chronic or of an
acute type. In the chronic type of infection in birds, the
leishmaniform, non-flagellate phase of the herpetomonad was the
more obvious, while in the more acute cases, a greater number of
flagellates was present. Monge (1914) made suggestions of a similar
import in regard to Lcishmania tropica in man in Peru. The parasites
are much more numerous in the internal organs of the experimental
birds than in the circulating blood, a feature common in kala-azar.
55 2
V. NATURAL HERPETOMONIASIS IN BIRDS
As before mentioned, in May, 1907, a short account of a
herpetomonad occurring in the blood of a pigeon was given by
Drs. Edm. and Et. Sergent. During their studies of the
Haemosporidia of birds, when working on the relations of
Haemoproteus columbae in pigeons and in the second host which
they discovered to be the Hippoboscid fly, Lynchia maura> they
found a herpetomonad in the blood of one of their Parisian pigeons.
The body of the herpetomonad was straight and drawn out,
measuring 17/z to 22/z by i‘5/*. The flagellum measured 19/1 to
35/1. No undulating membrane was present. The elongate
nucleus, measuring 5/1 to 7/* in length, was not as wide as the body,
and was situated 6 n to 7/1 from the posterior extremity of the body.
The large, spherical, heavily staining blepharoplast was 3/* to 5/1
in front of the nucleus. Care was taken by the authors to avoid
confusion with spermatozoa of the bird. Two excellent figures of
this herpetomonad were given. The parasite has not been observed
since. It is much to be regretted that an opportunity for following
further this interesting discovery did not arise.
The source of the herpetomonad is not known with certainty.
The bird containing the parasite had been inoculated intravenously
with part of the Berkefeld filtrate of an emulsion of ten Lynchia
maura previously fed on a pigeon infected with Haemoproteus
columbae . As the bird contracted the Haemoproteus infection, it
seems possible that this filtrate* also furnished the flagellate, or the
bird may have had a latent herpetomoniasis contracted direct from
insect hosts.
Considerable resemblances are presented by the account of this
presumed natural herpetomoniasis of the pigeon and the induced
herpetomoniasis in our birds—canary, sparrow and martin. The
flagellate form of the parasite is well marked in each host. The
free flagellum is distinct, the nucleus and blepharoplast have the
same staining reactions. The same parallel holds in the case of the
natural and induced herpetomoniases of mice recently described by
us. These parallel conditions suggest that these diseases induced
* Since this was written, Dr. Edmond Sergent has kindly informed us that so far as he was
aware, his Lynchia were not infected with herpetomonads. Our second hypothesis, then, may
be the correct one. We wish to thank Dr. Sergent for his courtesy and great interest.
553
by natural flagellates of invertebrates will, sooner or later, be found
in parallel under conditions not those of the laboratory. Some, in
fact, have already accrued (see Section VI).
VI. THE FLAGELLATE STAGE IN HERPETOMONIASIS
AND LEISHMANIASIS
Recently one of us pointed out in these Annals that the
flagellate form of Leishmania had been known for some time in
man, while it had been known for considerably longer in cultures.
Briefly, Escomel, in 1911, in Peru saw flagellate forms of
Leishntania tropica in man, and published about them later.
La Cava (1912) described similar forms of the same parasite in
Italy. In 1912 Splendore found elongating forms and a few
flagellate parasites in cases of dermo-mucosal leishmaniasis in
Brazil, while Monge (1914) found the herpetomonad stage of the
parasite of the same malady in Peru. The Leishmania cycle, then,
is that of the herpetomonad.
There was thus considerable evidence of the existence of
the flagellate stage of Leishmania in the vertebrate host at
the time when we suggested, as Patton has also done,
that canine kala-azar was really a herpetomoniasis due to
H. ctenocephaliy that leishmaniases were really arthropod-borne
herpetomoniases, and that it was ‘ likely that certain verte¬
brates . . . especially those that are insectivorous, may serve as
reservoirs for leishmaniases,' these conclusions having resulted from
our former series of experiments. We also stated that ‘ in areas
where leishmaniases are endemic, an examination should be made
of all insects and other invertebrates likely to come into contact with
men or dogs or rats and mice, in order to ascertain if these
invertebrates harbour herpetomonads. Preventive measures should
be directed against such invertebrates, especially arthropods,'
Additional support to the belief that leishmaniases are really
herpetomoniases has been received quite recently by the announce¬
ment that in September, 1915, Wenyon has found the flagellate
stage of Leishmania donovani in a dog subinoculated from
other dogs, the strain being derived from a man who died of
kala-azar contracted in Calcutta. He has also found variation in
554
the size of the Leishman-Donovan bodies in animals, as has been
previously noted by us in the non-flagellate stages of herpetomonads
introduced into vertebrates.
As we have shown elsewhere, flagellate as well as non-flagellate
forms of the parasites occurred in the internal organs of sticklebacks,
frogs, toads, snakes, mice and dogs, infected with Herpctomonas
jaculum, H. stratiomyiae , H. ctcnocephali , or H . pediculi. To this
list of hosts we can now add the canary, sparrow and martin, and
H. culicis increases the list of parasites. Comparing these experi¬
mental results with the finding of the flagellate stages of Leishmania
donovani and of L. tropica in dogs and in man, respectively, the
evidence that the parasites of leishmaniases are really herpetomonads
(or leptomonads, as some authors term them) seems thus conclusive.
It is not always easy to find flagellate forms of the various
herpetomonads that we have used. They are fragile, and are easily
broken, and disintegrate relatively soon after the death of the host.
Again, the flagellate stage in the development of the parasite is not
always present at the time of death. It is probably owing to these
two factors that the flagellate L. donovani has not been found direct
in man so far.
Recently one of us drew attention to the flagellate forms
described by Darling in connection with Histo plasma capsulatum.
It seems probable that the parasites in question may have been those
of a herpetomonad co-existing with rounded H. capsulatum ,
considered by many authorities to be a yeast. Up to the present, no
explanation of the significance of these flagellate forms has been
afforded, and before final classification of the organism can be made,
it will be necessary to elucidate whether these elements are
independent or have a connection with Histo plasma. Mixed
infections of herpetomonads and trypanosomes are capable of
co-existence, and this should also be remembered when so-called
4 herpetomonad phases of trypanosomes ’ are found.
The accumulation of evidence regarding the existence of a
flagellate stage in natural and induced herpetomoniasis in verte¬
brates shows the necessity for considering not part but the whole
of the life-history of an organism, and not only that, but the relation
of the parasite to the group to which it belongs. There is a line of
evolution common to each group, and in these cases, neither
555
Herpetomonas (. Leptomonas ), Leishmania nor Trypanosoma should
be considered as isolated units, but as flagellates belonging to the
Trypanosomidae. Much of the discussion that has arisen in latter
years would have been unnecessary were the organisms considered
from the broader, comparative standpoint.
VII. GENERAL CONCLUSIONS
The general conclusions that have resulted from our series of
experiments are as follows:—Under suitable conditions, insect
flagellates can be introduced into vertebrate hosts and produce
infection therein. In some cases, as in cold-blooded vertebrates, little
obvious ill-effect results; in others, as in mammals and birds, disease
is manifested and often ends in death. Similar infections are known
to occur naturally in some cases, for example, in mice and pigeons.
The organisms, such as herpetomonads, thus introduced, retain
their powers of development on the same lines as when they were
present in the insects. The morphological cycle of Leishmania is
like that of Herpetomonas. The various species of Leishmania are
probably insect herpetomonads long since introduced into man and
usually perpetuating the non-flagellate, relatively more resistant
form, though capable of assuming the flagellate, herpetomonad facies
in the internal organs of the vertebrate or in the invertebrate host.
Various vertebrates—fish, amphibia, reptiles, birds and
mammals—may serve as reservoirs of leishmaniases. The virus may
be very attenuated and so escape detection, or only be revealed by
the presence of the flagellate forms in cultures. It has also been
suggested by Stephens (1915) that each case of leishmaniasis in verte¬
brates arises de novo from the introduction of insect flagellates.
No insect flagellate can be considered to be quite innocuous to
vertebrates until it has been put to the test.
Leishmaniasis, which is a form of herpetomoniasis (lepto-
moniasis), is a flagellosis, as is also trypanosomiasis. Treatment of
leishmaniasis by intravenous injections of tartar emtic, as advocated
and practised recently by Vianna, Carini, di Cristina and Caronia,
Rogers and others, is sound biologically, as drugs containing arsenic
or antimony have proved efficacious in trypanosomiasis. It is thus
556
seen that the researches of the comparative morphologist are of the
greatest possible assistance to the medical man when founding a
basis for therapeutics.
VIII. SUMMARY
1. Herpetomoniasis can be induced in birds, for example,
canaries ( Serinus canarius ), sparrows ( Passer domesticus) and
martins ( Chelidon urbica :), by feeding them on insects containing
herpetomonads.
2. Herpetomonas culicis from Culex pipiens and H. jaculum
from Nepa cinerea have fatally infected birds when fed to them.
Both flagellate and non-flagellate herpetomonads have been found
in the internal organs of the infected host.
3. The cycle of the flagellates in the avian hosts resembled
morphologically that in the insects.
4. The disease induced may run an acute or a chronic course.
In the acute cases in our birds the flagellate form of the parasite was
the more obvious at death. In chronic cases, non-flagellate forms
of the parasite were more numerous.
5. Natural herpetomoniasis of a pigeon has been recorded by
Drs. Edm. and Et. Sergent in Algeria. This affords a parallel case
with the natural and induced herpetomoniasis in mice previously
recorded by us.
6. The flagellate stage of Leishmania donovani in vertebrates
is now known, and that of L. tropica in man has been known for
some time. The links completing the evidence that a Leishmania is
morphologically a Herpetomonas are thus complete. Leishmaniases
are really herpetomoniases (or leptomoniases) arising from herpeto¬
monads of certain invertebrates.
7. Members of all classes of vertebrates may be capable of acting
as reservoirs of herpetomoniasis, and the virus may exist in a very
attenuated condition and so be difficult of detection.
REFERENCES
Christophers, S. R. (1904). On a parasite found in persons suffering from Enlargement of
the Spleen in India. Sci. Mem. Goit. India , No. 11.
Darling, S. T. (1909). The Morphology of the Parasite (Histoplasma capsulatum) and the
Lesions of Histoplasmosis, a fatal disease of Tropical America. Journ. Exptl. Med., XI,
PP- 5 * 5 - 53 *- 5
Dutton, J. E., and Todd, J. L. (1903). First report of the Trypanosomiasis Expedition to
Sencgambia (1902). Part of Sect. VII. Flagellata in the Blood of a Mouse. Liverpool
Scb. Trop. Med.. Memoir XI. pp. 56-57.
557
Escomel, E. (1914). Leiahmania Flagelada cn cl Peru. La Cronica Medica [Lima], XXXI,
pp. 224-227.'
Fantham H. B. (1911). The Life-History of Trypanosoma gambiense and T. rkodesiense as
seen in Rats and Guinea-pigs. Proc. Roy. Soc., B, LXXXIII, pp. 212-227. 1 PI.
- (1912). Herpetomonas pedicvli, nov. spec., parasitic in the alimentary tract of Pediculus
vestimenti, the human body louse. Proc. Roy. Soc ., B, LXXXIV, pp. 505-517. 1 PI.
- (1915). Insect Flagellates and the Evolution of Disease, with remarks on the Importance
of Comparative Methods in the Study of Protozoology. Annals Trap. Med. Parasitol.,
IX, pp. 335-348.
Fantham, H. B., and Porter, A. (1913). Herpetomonas stratiomyiae , n.sp., a Flagellate Parasite
of the Flies, Stratiomyia chameleon and S. potamida , with remarks on the Biolog v of the
Hosts. Annals Trop. Med. and Parasitol., VII, pp. 609-620. 1 PI.
- (19*4). Some Insect Flagellates introduced into Vertebrates. Proc. Cambridge
Pbilosopb. Soc., XVIII, pp. 39-50. 1 PI.
-(1915)- Further Experimental Researches on Insect Flagellates introduced
into Vertebrates. Proc. Camb. Pbilosopb. Soc., XVIII, pp. 137-148.
-(191 ^). On the Natural Occurrence of Herpetomonads (Leptomonads) in
Mice. Parasitology, VIII, pp. 128-132.
La Cava, F. (1912). De la Leishmaniose des muqueuses et de la premiere decouverte de la
Leisbmania tropica HagclLe dans le corps humain. Bull. Soc. Path, Exot., V, pp. 808-812.
Layeran, A. (1915). Des Lacertiens peuvent-ils etre infectes par dcs Leishmania ? Bull.
Soc. Path. Exot ., VIII, pp. 104-109.
Laveran, A., and Franchini, G. (1914). Infections de Mammifercs par dcs flagclles
d’invert£br6s. Bull. Soc. Path. Exot., VII, pp. 605-612. 4 Figs.
Lindsay, J. W. (1914). American Forestal Leishmaniasis. Trans. Soc. Trop. Med. and
Hyg., VII, pp. 159-263.
Mesnil, F., Nicolle, M., and Remlinger, P. (1904). Sur le protozoairc du bouton d’Alcp.
C. R. Soc. Biol., LVII, pp. 167-169.
Monge, C. (1914). La Leishmaniasis del dermis cn el Peru. Espundia, Uta, Juccuya, Qccpo,
Tiacc-araoa. La Cronica Medica. [Lima], XXXI, pp. 231, 251, 288, 385.
Patton, W. S. (1907). Preliminary note on the life-cycle of a species of Herpetomonas found
in Culex pipiens. Brit. Med. Joum., July 13, 1907, pp. 78-80.
- (1908). The life-cycle of a species of Crithidia parasitic in the intestinal tract of Gerr is
fossarum. Arcb. /. Protistenkunde, XII, pp. 131-146. 1 PI.
-(1912]. The Morphology and Life History of Herpetomonas cultcis . Novv,
MacNeal and Torrey. Sci. Mem. Govt. India , No. 57.
Porter, A. (1909). The morphology and life-history of Critbidia gerridis, as found in the
British water-bug, Gerris paludum. Parasitology, II, pp. 348-366. 1 PI.
-(1909). The life-cycle of Herpetomonas jaculum (Leger), parasitic in the alimentary
tract of Nepa cinerea. Parasitology, II, pp. 367-391. 1 PL
- (*9*4)- The Morphology and Biology of Herpetomonas patellae , n.sp., parasitic in the
Limpet, Patella vulgata , together with Remarks on the Pathogenic Significance of
certain Flagellates found in Invertebrates. Parasitology, VII, pp. 322-329.
Rocha-Lima, H. da (1912). Beitrag zur Kenntnis der Blastomykosen. Centralbl. /. Baht.,
Abt. 1, Orig., LXVII, pp. 233-249. 1 PI.
Rogers, L. (1905). The Conditions affecting the Development of Flagellated Organisms
from Leishman Bodies and their bearing on the probable mode of Infection. Lancet ,
June 3, 1905, pp. 1484-1487. See also Lancet, July 23, 1904, p. 215.
Sergent, Edm. and Et. (1907). Etudes sur les Hcmatozoaires d’Oiseaux. Ann. Inst. Pasteur ,
XXI, pp. 251-280. 2 Pis. (For natural Hcrpctomonad in pigeon, see footnote, p. 270
and Plate VII.)
Sergent, Edm. and Et., Lemaire, G., and Senevet, G. (1914). Insccte transmcttcur ct
reservoir de virus du clou de Biskra. Hypoth^se et experiences prtliminaires. Bull.
Soc. Path. Exot., VII, pp. 577-579.
Stephens, J. W. W. (1915). The Mode of Transmission of some Tropical Diseases. Trans.
Liverpool Biol. Soc.. XXIX, pp. 3-19 (see p. 12).
Splendors, A. (1912). Lcishmaniosi con localizzazione nelle cavita mucose (nuova forma
clinica). Bull. Soc. Path. Exot., V, pp. 411-438. 2 Pis.
Wen von, C. M. (1915). Flagellate forms of Leisbmania donovant in the tissues of an experi¬
mentally infected dog. Journ. Trop. Med. and Hyg., XVIII, pp. 218-219.
55 »
EXPLANATION OF PLATE XXXIX
All figures were outlined with an Abbe-Zeiss camera lucida, using a
Zeiss ^ in. objective and compensating ocular 8. The magnification is
approximately 1,500 diameters.
Figs.
Figs.
Fig.
Figs.
Figs.
Figs.
Figs.
Fig.
Fig.
Fig.
Figs.
Fig.
Figs.
Figs.
Figs.
Figs.
Fig.
Figs.
Figs.
Fig.
Figs.
Fig.
Fig.
Fig.
Fig.
Fig.
Fig.
Fig.
Figs.
1-19. Hcrpctomonas jaculum , from the canary.
1-15. Non-flagellate forms of //. jaculum.
1, Small parasite. From circulating blood.
2, 9, Free forms. Heart smear.
3, 5. Lcishmaniform parasites. Bone-marrow.
4, 8. Non-flagellate forms. Lung.
6, 7. Parasites from kidney. Fig. 7 shows rounded blepharoplast and
karyosomatic nucleus.
10. Slightly curved parasite. Spleen.
11. Group of two pyriform parasites. Blood.
12. Group of small non-flagellate parasites around an erythrocyte.
Such groups are most uncommon. Blood.
13,14. Large parasites. Liver.
15. Elongating //. jaculum. Blood.
16, 17. Parasites showing root of flagellum. Fig. 16 shows a
karyosomatic nucleus. Lung.
18, 19. Flagellate forms of H. jaculum. Liver.
20-40. Hcrpctomonas culicis , from the sparrow and martin. Figs. 22,
23, 25, 30, 37, 39 from martin ; the rest from the sparrow.
20-26. Non-flagellate //. culicis. The parasite in Fig. 20 has the
blepharoplast superimposed on the nucleus. Figs. 20,21, from
blood ; Fig. 22, from bone-marrow ; Fig. 23, from lung ;
Fig. 24, from liver ; Figs. 25, 26, from heart smears.
26. Larger non-flagellate parasite.
27, 28. Two dividing parasites, from heart and bone-marrow
respectively.
29-40. Flagellate Hcrpctomonas culicis.
29. Very stout flagellate, probably about to divide. Heart.
30, 31. Flagellates from liver smears.
32. Parasite from kidney smear.
33. Flagellate with karyosomatic nucleus. Heart.
34. Flagellate from suprarenal capsule.
35. Parasite from heart, showing a few metachromatic granules.
36. Stout form from the lung. Round blepharoplast.
37. Flagellate folded on itself, as is frequent in life. Liver
38. Elongate flagellate. Liver.
39. 40. Dividing flagellates from the hearts of the martin and sparrow
respectively.
SS 9
ON THE ASSOCIATION OF
WARTHOG AND THE NKUFU TICK
(<ORNITHODORUS MOUBATA)
BY
LL. LLOYD
CHIEF ENTOMOLOGIST, N. RHODESIA
(Received for publication 12 November , 1915)
The following observation was made during searches for the
pupae of Glossina morsitans near Hargreaves, in the lower part of
the Luangwa Valley, N. Rhodesia. A native was sent into a
warthog burrow, in which the pupae are not infrequently found, to
scrape out the loose earth for examination. He was at once attacked
by a number of nkufu ticks with which the burrow was infested.
About thirty were removed from his clothing, and especially from
his head where they were actively feeding. Most of the ticks were
very young larvae which had not previously fed, some were nearly
full grown, and intermediate stages were represented. No adults
were seen.
The observation is of additional interest as regards the locality.
At the time that suggestions were being made as to possible carriers
of Trypanosoma rhodesiense , the nkufu tick, among other blood
suckers, fell under suspicion. It was pointed out, however, that it
was not generally distributed in the Luangwa Valley, the only
locality where it was known to occur being the compound of the
rubber plantation at Hargreaves (Neave). This plantation has now
been deserted for five years. The burrow in which the ticks were
found was on the opposite bank of the river, and the nearest village
was Mwapi, four miles away, where the tick is said not to occur.
560
Mr. L. C. Heath (N.C. at Mwengwa, N. Rhodesia) informed the
writer that he had removed a tick from a warthog which was
identified at Cambridge as Ornithodorus moubata. The warthog
is possibly of importance as a distributing agent for this pest, and
should not be overlooked in any prophylaxis against relapsing
fever.
561
ON ANAPLASMA-LIKE BODIES IN THE
BLOOD OF VERTEBRATES
BY
ANNIE PORTER, D.Sc. Lond., F.L.S.
BEIT MEMORIAL RESEARCH FELLOW, QUICK LABORATORY, CAMBRIDGE
(Received for publication 24 November, 1915)
Ten Text-figures
contents
Introduction . 561
Material and Methods . 561
Morphology ... ... ... . ... 562
Thi Nature of Anaplasmata . 564
Summary . 567
References . 568
INTRODUCTION
The study of vertebrate blood and its possible parasites is of
great importance, and many papers have been written during the
last ten years dealing with blood-parasites. Some of the papers
would have been much improved by a careful preliminary study of
the elements of apparently normal blood, or of blood under definite
pathological conditions. A number of the so-called parasites would
then have been found to be artefacts or reaction products, and not
organismal. Structures probably of this nature are the Anaplasmata
or ‘marginal points,* for which Theiler created the genus Anaplasma
in 1910, as he considered them to be organismal and the cause of
‘gall-sickness* in cattle in South Africa. Other investigators
consider Anaplasmata to be non-organismal, and to result from
haemolytic conditions of the blood.
MATERIAL AND METHODS
The material examined during these observations consists more
particularly of mice, canaries, swallows, martins, lizards, snakes,
frogs, toads and sticklebacks—that is, representatives of all the
great groups of the Vertebrata. The animals were used by me, in
collaboration with Dr. H. B. Fantham, on our researches into the
562
experimental introduction of insect flagellates—belonging to the
genera Herprtomonas and Crithidia —into different vertebrates.
The experiments resulted in induced herpetomoniasis in the verte¬
brates, often with pathogenic effects closely resembling those due to
leishmaniasis. The results of these experiments have been published
during 1914-15 in a series of memoirs listed in the References to this
paper. The presence of Anaplasmata was noted in the blood of the
experimental animals, many of which appeared to be anaemic. The
morphology, distribution and significance of the Anaplasmata seen
in the animals suffering from flagellosis will be considered.
Films of natural anaplasmosis in South African cattle have been
used for comparison, and examinations of blood films from obscure
cases of human anaemia have also been made.
MORPHOLOGY
Anaplasmata, when seen fresh, appear as small, rounded
granules or globules within the erythrocytes of the host. They show
no morphological differentiation. Usually they occur singly
(figs. 1, 2, 4), but forms apparently dividing and others simulating
rosette formation have been found, though such may be really
a gg re g a tions. When intra-vitam staining was employed, no
differentiated structures were observed within the globules.
Figs, i to 7. Anaplasma-like bodies from the blood of mammals, such ns young mice, x 1500.
Figs, i and 2. Bodies like Anaplasma marginale.
Fig. 4. Similar body like Anaplasma eentrale.
Figs. 3 and 5. Red corpuscles each containing two such bodies.
Fig. 6. Free form.
Fig. 7. Marginal point surrounded by a relatively clear area.
Stained preparations showed much the same features as fresh
ones. The structures always consisted of small, uniform, usually
spherical masses that stained intensely with chromatin stains, that
563
is, they were basophilic. Sometimes the bodies were oval in shape,
and only very rarely were they irregular. Very occasionally a some¬
what less densely staining portion of cytoplasm surrounding the
chromatinic bodies was observed (figs. 7, 10), but no marked exterior
limit to such an area or halo could be found, and the staining
appeared to be merely an idiosyncrasy of that portion of the host
cell. It may be mentioned that the Anaplasmata showed the same
type of structure whatever was the host from which they were
obtained—man, cattle, mice, birds, reptiles, amphibia or fish.
The Anaplasmata measure 0*3 fi to 2/1 in diameter, the latter size
being somewhat uncommon. Structures about 0*5/1 in diameter are
relatively numerous. Free forms may occur (fig. 6).
Sometimes two Anaplasmata are found apposed, a diplococcus-
like structure resulting. The two individuals may be equal (fig. 3) or
sub-equal (fig. 5), and appear to be the result of division.
Figs. 8 to 10. AnapUsma-likc bodies in the red blood corpuscles of cold-blooded vertebrates
such as snakes and frogs. X 1500.
Figs. 8 and 10. Red corpuscles of grass snake containing one and two Anaplasmata respectively.
Fig. 9. Red blood corpuscle of frog containing oval Anaplasroa.
Apparent multiplicative forms. Bodies that formerly would have
been considered to be of a multiplicative nature (figs. 3, 5) have been
observed by me in, a number of cases of anaemia due to maladies
such as herpetomoniasis. The process of formation of Anaplasmata
and of their pseudo-multiplication is as follows in nucleated red cells,
such as those of the snake.
Near one point of the periphery of the nucleus of the red cell, a
small bud appears. This tiny projection increases in size, and
gradually becomes somewhat spherical. The bud thus formed is
extruded and passes into the cytoplasm as a small, spherical body
(figs. 8, 9). This is the Anaplasma. Sometimes two buds appear
564
side by side, and pass outwards attached to one another. On other,
rarer occasions, more especially when the preparations have been
examined under dark-ground illumination, a small stream of five or
six buds is emitted from the nucleus, the result being a small rosette
of Anaplasmata in the cytoplasm of the host cell.
On a few rare occasions, the breaking up of large Anaplasmata
into smaller ones has been observed. The occurrence of more than
one Anaplasma in a corpuscle (fig. 10) may be thus explained, or
the bodies may have had separate origins. There is no special order
or sequence about this disruption, which resembles the fragmentation
seen in decaying nuclei of certain plant tissues. It is different from
the true division of any protozoon nucleus with which I am
acquainted.
The origin of the Anaplasmata in mammalian blood has only
been observed in a few instances. In such cases the host was a
young animal and anaemic, and also showed a certain number of
nucleated erythrocytes in its blood and in smears of organs made at
death. Anaplasma-forms were found in preparations of the spleen,
liver and bone-marrow, and similar bodies occurred free in the blood
plasma. There was no difference in morphology, whatever the
situation in which the structures were found.
I may mention that I have had the advantage of comparing my
preparations with blood films containing Anaplasmata, which films
came from Theiler’s laboratory in Pretoria.
THE NATURE OF ANAPLASMATA
The nature of the structures termed Anaplasmata has been the
subject of much controversy at different times, and very contradic¬
tory accounts of these bodies have been given by different workers.
The earlier workers on piroplasmosis, for example, Smith and
Kilborne (1893), noted small structures in the blood of cattle infected
with Texas fever. They described them as round coccus-like bodies,
o'2/i to 0 5 fji in diameter, which were usually situated at the
periphery of the corpuscle. Some larger granules were also seen.
These peripheral coccus-like bodies were considered to be probably
the early stages of Piroplasma bigeminum. They noted that the
peripheral bodies appeared in the blood as the number of blood
corpuscles began to fall, that is, under conditions of anaemia.
The foregoing conception of the nature of coccus-like bodies,
Anaplasmata or marginal points, continued in force till about 1910,
when Theiler, working in South Africa, put forward the suggestion
that the peripheral, coccus-like bodies of Smith and Kilborne were
organisms independent of Piroplasmata and producing a different
disease from the latter organisms. He considered the marginal
points, as the structures had been often termed, to be the true
excitants of gall-sickness in cattle, and created the genus Anaplasma
for them, two species, A . marginale and A. centrale being sub¬
sequently differentiated.
Since Theiler’s memoir appeared, a number of observations have
been published that show the presence of Anaplasmata in many
hosts, and in maladies not associated with gall-sickness or piro-
plasmosis. They have been found in young dogs and in marsupials
in Australia. Donkeys suffering from trypanosomiasis in the Sudan
show Anaplasmata, as do cats and rats suffering from the same
disease. Guinea-pigs, monkeys, lemurs, calves, sheep, goats,
horses, asses, pigs, rabbits, moles, mice, all have been reported as
containing Anaplasmata, many of these hosts apparently being
quite healthy. The bodies have been seen frequently in newly bom
animals. Certain authors, having observed coccus-like or
Anaplasma-like bodies in the blood of animals experimentally
infected with leishmaniasis, considered the Anaplasmata to be
possibly the initial stages of the Leishman-Donovan bodies.
It is very interesting to note that Sangiorgi (1915) found
Anaplasmata or marginal points in the splenic blood of a child from
Catania who was infected with infantile kala-azar, a disease
producing progressive anaemia. I, personally, have seen similar
bodies in the blood corpuscles of children suffering from obscure
forms of anaemia in England, and in the blood of a man returned
from the Tropics, who had perhaps previously had malaria.
Anaplasmata probably have more than one origin, as is seen from
the previous statements. Still another circumstance in which such
bodies are found in the host vertebrate has been revealed.by the
work of Dias and Aragao (1913-14). These workers conducted a
number of experiments on guinea-pigs, rabbits, dogs and cattle by
inoculating them with phenylhydrazine, nitrobenzol, pyrogallic
acid, saponin, phosphorus in oil emulsion and trypan blue. Nitro-
566
benzol, pyrogallic acid and phenylhydrazine produced structures
which were compared with preparations of natural Anaplasmata
obtained from Theiler. The staining reactions, size, morphology,
lack of structure and pseudo-division forms were the same as those
found in bovines naturally infected with anaplasmosis. The
injection of haemolytic substances thus produced structures of the
same nature as Anaplasmata. The authors conclude that
Anaplasma is not a protozoon, but is a product of the degeneration
of the red cells due to the action of the haemolytic substances
introduced into the host. The anaplasmosis of Brazil is considered
to be really a piroplasmosis, in which the parasites remain chiefly in
the internal organs of the host. The presence of Anaplasmata in
animals infected with worms is considered to be due to the
haemolytic action of the toxines produced by the helminthes, while
their occurrence in newly-born animals is ascribed to the activity of
their haematopoietic organs. Laveran and Franchini (1914) have
confirmed the production of Anaplasma-like bodies by the use of
phenylhydrazine.
Early in 1915, a paper by Veglia appeared containing an account
of the results obtained by him in culturing Anaplasma marginal
using various media. He found that the number of corpuscles
showing Anaplasmata increased greatly in the cultures. At lower
temperatures, the round form predominated. I.ater, the parasites
became larger and assumed a somewhat triangular or quadrangular
shape. Schizogony into three, four and eight is believed by him to
occur. Diplococcoid forms were also observed. A series of figures
showing the great increase in the numbers of infected corpuscles is
given. Regarding both the increase in numbers of the Anaplasmata
and of the infected corpuscles, I would suggest that another interpre¬
tation may be given. It seems to me that in these cases haemolytic
substances are produced in the culture medium from chemical changes
in some of the red corpuscles. The quantity of such substances
increases with the progress of the experiment, and acting on the
remaining red corpuscles, a number of Anaplasmata are produced;
the numbers of infected erythrocytes would, consequently, increase for
some time. As there may have been an increase in the quantity of
haemolytic substances in the blood of an animal suffering from
masked piroplasmosis, Veglia’s conclusion that the progress of his
567
cultures was on the same lines as that of natural anaplasmosis is
explicable.
It must be mentioned that Franchini and Mantovani (1915) found
in a culture of rat’s blood on Novy’s medium mixed with glucose,
a number of small bodies presenting the appearance of Anaplasma
in stained preparations. It is possible that the presence of
haemolytic substances in the culture resulted in the production of
these bodies.
It is interesting to note with regard to the origin of some
Anaplasmata, that over twenty years ago, Smith and Kilborne, with
remarkable foresight, suggested that some of the bodies now termed
Anaplasma were ‘ probably remnants of the nucleus of the ancestor
of the [red] corpuscle—the haematoblast.’
My own experiences suggest that Anaplasmata may be of nuclear
origin, and may be the results of anaemia induced in the host by
such diseases as herpetomoniasis.
If, however, Anaplasma be considered organismal, it affords
an interesting example of what is, perhaps, a phylogenetic and
recapitulative type of primitive Protozoon. Anaplasma might also
represent an organism which has been secondarily reduced in size
and structure.
SUMMARY
Anaplasmata may occur in healthy and in anaemic vertebrate
blood. The structures, also called marginal points and peripheral
coccus-like bodies, are probably of diverse origin. It is doubtful if
they are organismal in nature.
Anaplasmata have been found by me in warm and cold-blooded
vertebrates, wherein conditions such as herpetomoniasis and
anaemia occurred. Some of the bodies originate from the nucleus
of the erythrocyte or erythroblast, under the influence of haemolysis.
The Anaplasma-like bodies were basophilic, apparently
composed of chromatin or of a substance giving a similar staining
reaction, and were homogeneous in structure. They varied from
0*3 fi to 2*0 fi in diameter, often being about 0‘5/i. Binary and
multiple forms, which might be interpreted as phases of division,
were seen.
568
REFERENCES
Dias, E. C., and AragXo, H. de B. (1914). Pcsquizas sobre a naturcza dos Anaplasmas. Mem.
Inst. Ostvaldo Cruz, Vol. VI, pp. 231-249. 2 plates. (See also Brazil Medico , April 22,
I 9 , 3 *)
Fantham, II. B., and Porter, A. (1914-15). Some Insect Flagellates introduced into Verte¬
brates. Proc. Camb. Pbilosopb. Soc., Vol. XVIII, pp. 39-50. 1 plate. Also Cambridge
Univ. Reporter, Vol. XLV, p. 323 (Dec. 1, 1914).
-(1915). Further Experimental Researches on Insect Flagellates introduced
into Vertebrates. Proc. Camb. Pbilosopb. Soc., Vol. XVIII, pp. 137-148. Also Cambridge
Univ. Reporter, Vol. XLV, p. 931 (May 25, 1915).
-(1915). On the Natural Occurrence of Herpetomonads (Lcptomonads) in
Mice. Parasitology , Vol. VIII, pp. 128-132.
-( , 9 i 5). Some Experimental Researches on Induced Herpetomoniasis in
Birds. Annals Prop. Med. Parasitol., Vol. IX, pp. 543-558. 1 plate. Also
Cambridge Univ. Reporter , Vol, XLVI, p. 262 (Nov. 30, 1915).
Franchini, G., and Mantovani, M. (1915). Infection experimental du rat et de la souris
*par Hcrpetomonas muscae domesticae. Bull Soc. Path. Exot., Vol. VIII, pp. 109-m.
Lavf.ran, A., and Franchini, G. (1914). Contribution a l’ctude des ‘marginal points’ des
hematies de mammif^res. Bull. Soc. Path. Exot., Vol. VII, pp. 580-584.
Sangiorgi, G. (1915). Rcperto di 4 corpi anaplasmasimili nel sangue dell ’ uomo e degli animali.
Patbologica , Vol. VII, pp. 27-29.
Smith, T., and Kilborne, F. L. (1893). Investigations into the nature, causation and
prevention of Southern Cattle Fever. 8 tb and 9 tb Annual Report, Bureau of Animal
Industry , Washington, pp. 177-302. 10 plates.
Tiieiler, A. (1910). Anaplasma marginale (gen. et spec. nov.). The marginal points in the
blood of cattle suffering from a specific disease. Report of Govt. Vet. Bacteriologist,
Transvaal Dept, of Agric., for 1908-09, pp. 7-64.
-(1910). Anaplasma marginale (genus nov. et species nov.). Un nouveau protozoairc
du b£tail. Bull. Soc. Path. Exot., Vol. Ill, pp. 135-137-
Veglia, F. (1915). Coltura dell’ Anaplasma marginale in Vitro. Nota preliminare. Giorn.
R. Accad. Med. di Torino , Vol. LXXVIII, pp. 33-39.
INDEX
INDEX
PAGE
Index ok Authors . iii
General Index . iii
Index of Species and Varieties new to Science . x
INDEX OF AUTHORS
PACE
Bayon, H.I, 535
Biacklock, B. ; and Yorke, \V., 349, 363,
383, 4 X 3, 425
Breinl, A.213, 285
Breinl, A.; and Priestley, H. 495
Carter, H. F. 173
Davey, J. B. 421
Fantham, H. B.335, 391
Fantham, H. B.; and Porter, A. 543
Fletcher, VV.; and Fraser, H. 381
Fraser, H. ; and Fletcher, W. 381
Hirsch, E. F.; and Ward, H. B. 109
Lloyd, L. 559
Macfie, J. W. S. 435 . 457 . 5 ° 7 > 5 2 7
PAGE
Porter, A. 561
Porter, A.; and Fantham, H. B. 543
Priestley, H. 233
Priestley, H. ; and Breinl, A. 495
Scott, H. H. 239
Schwetz, J.163, 513
Seidelin, H. 197
Stephens, J. W. W.169, 429, 539
Stephens, J. W. W. ; and Stott, W.... 201
Stott, W. ; and Stephens, J. W. W.... 201
Ward, H. B. ; and Hirsch, E. F. 109
Yorke, W. ; and Biacklock, B., 349, 363,
383, 413, 425
Young, W. J. 91
GENERAL INDEX
PAGE
Accra, West Africa, Babesiasis in. 457
„ „ „ Trypanosomiasis
in . 457
Alimentary system of Paragonimus ... 123
Amblyomma variegatum . 419
Amoebic dysentery in monkey ...507, 508
Anaplasma-like bodies in blood of
Vertebrates.561, 567
Anaplasma marginale .233, 565, 566
Anaplasmata, Nature of.564, 567
Animal parasites of domestic stock in
Sierra Leone . 413
Ankylostoma braziliettse . 427
Ankylostoma caninum .425, 427
,, „ Dimensions of... 426
,, „ Mouth of . 425
„ „ Posterior ray of
bursa . 426
,, ceylanicum .425, 427
„ „ Dimensions of 426
,, „ Mouth of . 425
„ „ Posterior ray of
bursa . 426
page
Ankylostomiasis in dogs in Sierra
Leone.425, 427
Aponomma decorosum . 236
Arneth counts in health in malaria and
yellow fever com-
P are d . 45 2 > 453
.. „ in healthy Europeans in
West Africa, 437, 438,
456
,, „ in healthy natives in
West Africa.439, 440
„ ,, in malaria ...435,442,456
,, ,, in natives of New
Guinea ...495, 504, 505
„ „ in yellow fever ...435, 449,
456
„ „ Significance of changes
observed in.454, 456
Arneth index in natives in New
Guinea, 495, 500, 503,
504, 505
,, ,, Personal variations in
determining healthy 436
IV
PAGE
Artificial cultivation of Hansen’s
bacillus...23, 381, 535
Atypical cases of enteric fever . 266
Babesia bigemina . 460
,, 1 an is . 462
Babesiasis at Accra, West Africa . 457
„ of cattle and sheep in West
Africa. 457
Bacillus leprae in relation to diphthe¬
roids . 381
Bacillus of leprosy, Cultivation of. 381
Bacillus typhosus in the gall bladder, 254,
z6 5
,, ,, Peculiarities of . 262
Bacteriological investigations of enteric
fever. 253
Baobab as pupal habitats of Glossina
palpalis .35H 359
Bayon, H. Leprosy : A perspective
of the results of experimental study
of the disease . 1
Bavon, H. The Artificial Cultivation
of Hansen’s ‘ Bacillus’ . 535
Bile, Cultivation of bacilli from, in
enteric fever. 253
Bionomics of Glossina palpalis in Sierra
Leone .349, 361
Birds, Induced herpetomoniasis in ... 543
Blacklock, B., and Yorkc, W. Anky¬
lostomiasis in dogs in Sierra Leone... 425
Blacklock, B., and Yorke, W. Food
of Glossina palpalis in the Cape
Lighthouse Peninsula, Sierra Leone 363
Blacklock, B., and Yorke, W. Notes
on certain animal parasites of
domestic stock in Sierra Leone. 413
Blacklock, B., and Yorke, W. Notes
on the bionomics of Glossina
palpalis in Sierra Leone, with
special reference to its pupal
habitats . 349
Blacklock, B., and Yorke, W. The
reservoir of the human trypanosome
in Sierra Leone . 383
Blackwater fever associated
with quartan malaria. 429
„ ,, Blood in . 430
„ „ Chart of . 433
„ „ Conclusions on ... 432
,, „ History of case ... 429
„ „ Post-mortem. 431
PAGE
Blackwater fever associated
with quartan malaria,
Record of quinine
in . 430
•> » Urine in . 431
Blackwater fever, Relationship of
quinine to . 201
Blood, Arneth counts in.435, 495
Boophilus australis . 419
Breinl, A. Gangosa in New Guinea
and its etiology . 215
Breinl, A. On the occurrence aftd
prevalence of diseases in British
New Guinea. 285
Breinl, A., and Priestley, H. Differen¬
tial counts and the neutrophile
blood-picture of natives—adults and
children—of New Guinea. 495
British New Guinea, Contracting sore in 309
„ ,, „ Curious fevers in 297
„ „ „ Diseases in. 285
,, „ „ Filariasis in. 290
» » „ Gangosa in. 314
„ „ ,, Juxta-articular
nodules in ... 294
» » „ Leprosy in. 293
„ „ „ Malaria in . 288
„ „ „ Peculiar disease
in, charac¬
terised by
bone lesions... 300
,, „ „ Skin diseases in 314
„ „ „ Tropical slough¬
ing phage-
daena in. 306
, ,, „ Tropical ulcers
in .304, 306
„ „ „ Ulcus interdigi-
tale destruens
r in... 313
„ „ „ Venereal diseases
in. 29c;
» i> » Yaws in . 298
Canary, Herpetomonads introduced
into...545> 54-
Canine babesiasis at Accra . 462
Carriers, Unrecognised, in enteric
fever .239, 231
Carter, H. F. On some previously
undescribed Tabanidae from
Africa . 173
X
V
PAGE
Cattle, Epidemics among, in Sierra
Leone .414, 420
Chrysops stigmaticalis . 173
Comparative methods in Protozoology 335
Contracting sore in British New
Guinea . 309
Crithidia , Introduced into vertebrates, 342,
34 6
Cryptococcus mutilans , n.sp. 224
Culcx duttoni . 167
„ invidiosus . 167
„ tigripes . 167
,, univittatus . 167
Culiciomyia nebulosa . 168
Cultivation of leprosy bacillus, 23, 381, 535
Curious fevers in British New Guinea 297
Cuticular spines of Paragonimus.. .11% y 148
Davey, J. B. The etiology of juxta-
articular subcutaneous nodules. 421
Diatomineura acthiopica . 173
Differential blood counts in native
adults and children in New Guinea, 495,
496,505
Diphtheroids from leprous nodules ... 535
,, in relation to Bacillus
leprae . 381
Diseases in British New Guinea . 285
Dogs, Ankylostomiasis in, in Sierra
Leone .425, 427
Domestic stock, Animal parasites of*
in Sierra Leone . 413
Eloeis guineensis .350, 361
Enteric fever, Atypical cases of . 266
„ „ Bacteriological investi¬
gations of . 253
„ „ Cultivations from the
bile in. 253
„ „ Flies in relation to. 248
„ „ in Kingston, Jamaica... 239
„ „ Legislative recommen¬
dations in Kingston 268
„ „ Milk in relation to. 247
,, „ No post-mortem
evidence of, but posi¬
tive result on cultiva¬
tion of bile .258, 272
„ „ Personal contact in ... 267
„ „ Post-mortem signs of,
and positive result
on cultivation of
bile.256, 272
PAGE
Enteric Fever, Sewage disposal in
relation to . 249
„ „ Water supplies in
relation to . 245
Entamoeba cercopitbeci , n.sp. 510
Epimys rattusy trypanosome in . 527
Eretmapodites chrysogaster . 166
Etiology of juxta-articular sub¬
cutaneous nodules.421, 423, 424
Euphorbia , Herpetomonads in . 341
Evolution of disease, Insect flagellates
in relation to . 335
Excretory system of Paragonimus . 128
Fantham, fl. B. Insect flagellates and
the evolution of disease, with
remarks on the importance of
comparative methods in the study
of Protozoology . 335
Fantham, H. B. Spirochaeta bronchia -
liSy Castellani, 1907, together with
remarks on the spirochaetes of the
human mouth. 391
Fantham, H. B., and Porter, A. Some
experimental researches on induced
herpetomoniasis in birds . 543
Filariasis in British New Guinea. 290
Filaria volvulus . 423
Fletcher, W., and Fraser, H. The
cultivation of the leprosy bacillus... 381
Flies and enteric fever . 248
Food of Glossina palpalis in Sierra
Leone . 363
Fraser, H., and Fletcher, W. The
cultivation of the leprosy bacillus... 381
Gangosa, Clinical account of. 215
„ Description of cases of. 216
„ Etiology of. 221
„ in British New Guinea, 213, 314
Genus Paragonimus . 146
Glossina palpalis , Bionomics of ...349, 361
„ „ Difficulties encountered
in examining a
region for .
. 520
yy
yy
Dissection of
. 363
yy
yy
Distribution
in
Lomami .
. 5*3
yy
yy
Experimental
solu-
tions as food, 375, 379
yy
yy
Food of .
...363,378
yy
yy
Haemoglobin
as
food.
••• 373 , 379
VI
PAGE
Glossina palpalis , Pupal habitats in
relation to water, 352, 361
„ ,, Relative distribution
of males and
females . 525
,, „ Vegetable juices as
food.368, 378, 379
,, ,, Vertebrate blood as
food.....364, 378
,, „ Wild, in Sierra Leone 415
Goats, Trypanosomes in.465, 466
Granule stage of Spirochaetes ...399, 401
Haematopota angustifrons , n.sp. 185
„ corsoni , n.sp. 190
„ exiguicornuta, n.sp. 188
„ pinguicornis , n.sp. 182
„ scutellaris . 173
„ tkeobaldi , n.sp. 179
,, transvaalensis, n.sp. 176
„ West African species,
Table of. 193
Hacmocystozoon brasiliensc . 339
Haemoglobinuria, Duration of. 539
Hansen’s bacillus.1, 18
Hansen’s bacillus, Artificial cultivation
of.V.23. 535
Herpetomonad stage of Leishmania,
Existence in man 339, 55 3
„ stage of Leishmania,
Significance of 337,346, 553
Herpetomonads in plants .341, 346
„ Natural, in mice, 339, 340,
34 6
» „ ,, birds. 552
Herpetomonas introduced into verte¬
brates .342, 346
Herpetomonas rulicis introduced into
birds .546, 550
„ jaculum introduced into
birds .545, 549
Herpctomoniasis, Anaplasma-like
bodies in . 562
„ Induced, in birds... 543
,, Natural, in birds... 552
Hirsch, E. F., and Ward, H. B. The *
species of Paragonimus and their
differentiation. 109
Histoplasvia capsulatum , Flagellate
stages of ...339, 554
Human trypanosome in an ox in Sierra
Leone ...385, 389
PACE
Human trypanosome, Reservoir of, in
Sierra Leone 383
Hump-backed cattle,Trypanosomes in 465
Insect flagellates and the evolution of
disease . 335
„ „ introduced into
vertebrates... 342,543
Juxta-articular nodules in British New
Guinea . 294
„ subcutaneous nodules,
Etiology 0^421,423,424
„ subcutaneous nodules
in Nyasaland . 421
Kabinda (Lomami), Belgian Congo,
Mosquitos of . 163
Kala-azar, Preventive measures against, 345,
346
Kedrowsky’s bacillus.26, 382, 536
Kingston, Jamaica, Enteric fever in... 239
Koch’s bodies .235, 419
Kg or salvarsan-copper . 197
Lake Boya, Distribution of Glossina
palpalis around . 516
Legislative recommendations for
enteric fever in Kingston . 268
Leishmania, Herpetomonad stage,
existence in man...339, 553
„ Herpetomonad stage,
significance of..337,346,553
Leprosy Bacillus, Cultivation of, 23, 381,
535
Clinical nosology.4, 78
Communicability .55, 79
Diagnosis, differential. 67
Diffuse or hyperaemic. 8
Experimental study of. I
Experimental transmission to
animals .31, 78
Geographical distribution ... 2
Historical . 1
In British New Guinea.2, 93
Lupoid or serpiginous . 6
Maculo-anaesthetic. 5
Microscopical features of
lesions .9, 78
Mixed . 6
Nodular. 5
Prevention.73, 79
Prognosis, early . 64
Segregation . 74
Serology of .47, 79
Vll
PACE
T ,eprosy, Tuberculin and similar tests in 52
„ Treatment.70, 79
Leptomonas, Generic characters . 340
Lloyd, LI. On the Association of
Warthog and the Nkufu Tick
(Ornithodorus moubata) . 559
Lomami, Belgian Congo, Distribution
of Glossina palpalis in. 513
Macfie, J. YV. S. Babesiasis and try¬
panosomiasis at Accra, Gold Coast,
West Africa. 457
Macfie, J. W. S. Nuclear variations
of the neutrophile leucocytes
(Arneth counts) in malaria and
yellow fever. 435
Malaria, Arneth counts in...435, 442, 456
,, in British New Guinea . 288
,, parasite, peculiar morpho¬
logical appearances. 169
Man as a reservoir of trypanosomiasis
in Sierra Leone .384, 388, 389
Mangrove swamps and Glossina pal -
polis .355.361
Mansonioides uniformis . 167
Martin, Herpetomonads introduced
into.547, 548
Mare, Small monomorphic trypano¬
some in . 480
Metabolism of white races in the
tropics . 91
Milk in relation to enteric fever . 247
Monkey, Amoebic dysentery in . 507
„ Spirochaetes in large intes¬
tine and rectum of ...507, 509
Morphological peculiarities of a
malarial parasite. 169
Mosquitos of Kabinda, Belgian Congo 163
„ Papuan, list of. 317
Mules, Trypanosomiasis in, resembling
dourine. 486
Mus decumanus, New piroplasm in ... 462
Mycobacterium leprae . 1
„ „ Artificial culti¬
vation, 23,77, 381,
535
„ „ Morphology ... 18
„ „ Staining. 22
Myzomyia costalis . 166
Neutrophile leucocytes in native adults
and children in New
Guinea.495, 504, 505
PACK
Neutrophile leucocytes, Nuclear varia¬
tions of . 433
New Guinea, Differential blood counts
in native adults and
children.495,496, 505
Gangosa in . 213
Neutrophile leucocytes
in native adults and
children.495, 504, 505
.-. 559
Nocardia carougeaui . 424
Nuttallia decumani , n.sp.462, 464
Ochlerotatus (Protomacleayd) albozrn-
tralis . 168
Oil palms in relation to pupae of
Glossina palpalis .352, 358
,, Removal of lower petioles
in relation to Glossina
breeding grounds, 353, 360,
361
vu 11 f 11 tirb
Ornithodorus moubata . 559
Ox, Human trypanosome in, in Sierra
Leone.385, 386, 389
Paragonimus, Alimentary system . 123
,, Cuticular spines.118,148
,, Excretory system . 128
„ Genus discussed . 146
„ Historical. no
,, Reproductive system ... 129
Species of, and their
differentiation ...109, 116
„ Structure of. 116
Paragonimus kellicotti ... 115
,, kellicotti , Cuticular
spines . 120
„ ringeri . 117
„ ringer 1, Cuticular
spines . 121
,, westermanni . 117
„ westermanni , Cuticular
spines . 121
Papuan mosquitos, List of . 317
Peculiar disease characterised by bone
lesions in British New Guinea. 300
Personal contact in enteric fever. 267
Piroplasma bigeminum .414, 418, 420
Plasmodium tenue . 169
Polymorphic trypanosome in mules at
Accra, Measurements of. 488
Porter, A. On Anaplasma-like bodies
in the blood of vertebrates . 561
Porter, A.; and Fantham, H. B.
Some experimental researches on
induced herpetomoniasis in birds... 543
Priestley, H. Theileria tachyglossi,
n.sp., a blood parasite of Tachyglossus
aculeatus . 233
Priestley, H., and Breinl, A. Differen¬
tial counts and the neutrophile
blood-picture of natives—adults and
children—of New Guinea. 495
Protein metabolism of white races in
tropics . 91
Protozoology, Comparative methods
i n . v . : . 335
Pupal habitats of Glossina palpalis in
relation to water, 352,
361
„ „ Glossina palpalis in
Sierra Leone, 349, 361
Quartan malaria associated with black-
water fever . 429
Quinine and rigor, Relationship be¬
tween .207, 209
„ in relation toblackwater fever 201
Rat leprosy in relation to the human
disease .40, 78
„ „ Attempts to cultivate the
bacillus of . 42
Reproductive system of Paragonimus. .. 129
Reservoir of human trypanosome in
Sierra Leone.383, 390
Rhinopharyngitis mutilans . 213
River Kekcy, Distribution of Glossina
palpalis around. 517
„ Lomami, Distribution of Glos¬
sina palpalis around . 518
Salvarsan-copper in trypanosomiasis... 197
Schwetz, J. Preliminary notes on the
mosquitos of Kabinda (Lomami),
Belgian Congo . 163
Scott, H. H. An investigation into
the causes of the prevalence of
enteric fever in Kingston, Jamaica ;
with special reference to the question
of unrecognised carriers. 239
Seidelin, H. Experiments with
salvarsan-copper in trypanosomiasis 197
Sewage disposal and enteric fever. 249
Sheep, trypanosomes in .465, 466
Sierra Leone, Animal parasites of
domestic stock in . 413
Sierra Leone, Ankylostomiasis in dogs, 425,
427
„ „ Cape Lighthouse Penin¬
sula . 363
„ Glossina palpalis in, 349, 361,
363
„ „ Reservoir of human try¬
panosome in.383, 390
Skin diseases in British New Guinea... 314
Sparrow, Herpetomonads introduced
into .546, 548
Species of Paragonimus . 116
Spirochaeta bronckialis . 391
„ „ Geographical
distribution.. 407
„ „ Granule
phase ...399,408
„ „ Mode of in¬
fection, 404, 408
„ „ Morphology 395,
408
„ „ Morphological
variations ... 396
„ „ Movements 394,
408
„ „ Pathogenicity 405
„ „ Treatment for 407
Spirochaeta buccalis .402, 409
„ dentium 402, 409
„ curygyrata . 510
Spirochaetes, Granule stage of. 401
„ like S. curygyrata in gut
of monkey.507, 510
„ of the human mouth... 391
„ of the human mouth,
morphology. 402
„ of the human mouth,
contrasted with S.
bronchialis . 404
Stegomyia africana . 167
„ apicoargentea . 167
„ fasciata . 167
„ power i . 167
„ simpsoni . 167
Stephens, J. W. W. On the peculiar
morphological appearances of a
malaria parasite . 169
Stephens, J. W. W. Studies in black-
water fever. IV—Note on a case
of quartan malaria associated with
blackwater fever . 429
\x
PAGE
Stephens, J. \V. \V. Studies in black-
water fever. V—Duration of
Haemoglobinuria . 539
Stephens, J. W. W., and Stott, W.
Studies in blackwater fever.
Ill—The relationship of quinine
to blackwater . 201
Stott, W., and Stephens, J. W. W.
Studies in blackwater fever.
Ill—The relationship of quinine
to blackwater . 201
Straight-backed cattle, trypanosomes
in.465, 466
Structure of Paragonimus . 116
Tabanidae from Africa . 173
Tabanus atrimanus . 173
„ ditaeniatus . 173
,, fuscipes var. oculipilus , n. var. 175
„ insignis . 173
„ sericeiventris . 173
„ taeniola . 173
,, triquetrornatus , n.sp. 173
T achy gloss us aculeatus , Blood parasite
° f .. 2 33
Taentorhynchus aurites . 167
„ cristatus . 167
Theileria mutans .419, 460
„ mutans in cattle in Sierra
Leone.419, 420
„ parva . 419
„ tachyglossi , n.sp. 233
„ tachyglossi , Morphology of... 234
Toxor/yynchites brevipalpis . 166
Tropical ulcers in British New Guinea 304,
306
„ sloughing phagedaena in
British New Guinea . 306
Tropics, White races in, Protein meta¬
bolism of . 91
Trypanosoma congolense in animals at
Accra, 464, 465, 472, 477,
478
„ congolense in cattle 414, 415,
420
congolense , Measurements
° f .•..473
congolense var. equinum
(n. var.). 486
eburneense . 531
equi, Parasites resembling,
in mules in Accra, 490, 491
PAGE
Trypanosoma gambiettse in an ox in
Sierra Leone .386, 389
gambiense in cattle 414, 415,
420
longocaudense . 530
„ pccaudi in animals at
Accra, 464, 465, 466, 478,
486
„ pecaudiy Measurements of 468
„ vivax in animals at
Accra...464, 465, 470, 478
„ vivax in cattle 414, 415, 420
„ vivaxy Measurements of 470
Trypanosome of the black rat (Epimys
rati us) 527
„ „ „ Measure¬
ments... 5 29, 530
„ „ „ Morphology.. 528
Trypanosome, small monomorphic, in
blood of a mare. 480
„ „ „ Measure¬
ments of 482,484
Trypanosomes in goats .465, 466
„ in hump-backed cattle 465
„ in sheep .465, 466
„ in straight-backed
cattle . 466
„ So-called herpeto-
monad stages of.341,
345.346
Trypanosomiasis at Accra, \\ est Africa, 457
464
„ in dogs in Accra ... 477
„ in equincs . 478
„ Resembling dourine,
in mules . 486
„ Salvarsan-copper in 197
Ulcus interdigitale destruens in British
New Guinea. 313
Unrecognised carriers in enteric fever 239
Urine of persons in tropics,
Ammonia in 97
„ „ „ Chlorides... 99
„ „ „ Creatine ... 97
„ „ „ Creatinine .. 97
„ „ „ Phosphoric
acid . 99
,, „ „ Purin bases 99
„ „ „ Specific
gravity of 95
„ „ „ Sulphur ... 99
TACiL
Yellow fever, Arncth counts in
PACE
Urine ol persons in tropics,Total
nitrogen of 95
„ „ Urea in. 97
„ „ „ Uric acid ... 98
„ ,, „ Volume of... 95
Venereal diseases in British New
Guinea. 299
Vertebrates, Anaplasma-like bodies in
blood of.561, 567
Ward, II. B., and llirsch, E. E. The "
species of Paragonimus and their
differentiation. 109
Warthog and Nkufu tick . 559
Water supplies in relation to enteric
fever. 245
West African species of llaematopota ,
Tabic of . 193
White races in tropics, Protein meta¬
bolism of . 91
Yaws in British New Guinea . 298
„ in relation to juxta-articular
nodules.422, 423
435 ,
449.456
Yorke, \Y\, and Blacklock, B. Anky¬
lostomiasis in dogs in Sierra Leone... 42;
Yorke, \Y\, and Blacklock, B. Food
of Glossina palpalis in the Cape
Lighthouse Peninsula, Sierra Leone 363
Yorke, W., and Blacklock, B. Notes
on certain animal parasites of
domestic stock in Sierra Leone.413
Y'orke, W., and Blacklock, B. Notes
on the bionomics of Glossina palpalis
in Sierra Leone, with special
reference to its pupal habitats. 349
Yorke, W., and Blacklock, B. The
reservoir of the human trypanosome
in Sierra Leone . 383
Young, \Y\ J. The metabolism of
white races living in the tropics.
I—The protein metabolism . 91
INDEX OF SPECIES AND VARIETIES NEW TO SCIENCE
PACE
Cryptococcus mutilans . 234
Entamoeba ccrcopitheci .510, 512
flaematopota angustifrons . 185
„ corsoni . 190
„ exiguicornuta . 188
„ pinguicornis . 182
,, theobaldi . 179
PAGl
Hacmatopota transzaalensis . 176
Suttallia decumani . 462
Tabanus juscipes , var. oculipilus , n. var. 175
,, triquetrornatus . 173
Theileria taehyglossi .233, 236
Trypanosoma congolense , var. equinum ,
n. var. 48^
Live Music
Archive
Librivox
Free Audio
Metropolitan Museum
Cleveland
Museum of Art
Internet
Arcade
Console Living Room
Open Library
American
Libraries
TV News
Understanding
9/11